Fluorescence resonance energy transfer mapping of the fourth of six nucleotide-binding sites of chloroplast coupling factor 1.
AuthorsShapiro AB, Gibson KD, Scheraga HA, McCarty RE
JournalJ Biol Chem
PubMed ID1832671
Equilibrium dialysis measurements of adenine nucleotide binding to chloroplast coupling factor 1 suggest that the enzyme has six binding sites for ADP, adenylyl-beta,gamma-imidodiphosphate (AMP-PNP), and 2'(3')-O-2,4,6-trinitrophenyl-ATP (TNP-ATP). High affinity binding at all six sites requires the divalent cation, Mg2+. Three of the nucleotide-binding sites, sites 1, 2, and 3, have ... More
Site-site interaction on mitochondrial F1-ATPase. Functional symmetry of the high-affinity nucleotide binding sites.
AuthorsTiedge H, Schäfer G
JournalBiol Chem Hoppe Seyler
PubMed ID2876715
Interactions between the high affinity binding sites on mitochondrial F1 were analysed by combined use of the nucleotide analogues 3'-O-(1-naphthoyl)-ADP (N-ADP) and 2'-3'-O-(2,4,6-trinitrophenyl)-ADP (TNP-ADP). The binding behaviour of F1 with respect to these ligands was studied by measuring the fluorescence of F1 and of TNP-ADP and the fluorescence anisotropy of ... More
Mitochondrial ATP synthase. cDNA cloning, amino acid sequence, overexpression, and properties of the rat liver alpha subunit.
AuthorsLee JH, Garboczi DN, Thomas PJ, Pedersen PL
JournalJ Biol Chem
PubMed ID2137825
'The predicted amino acid sequence of the alpha subunit of the rat liver mitochondrial ATP synthase has been obtained by sequencing a cDNA for the alpha subunit. Analysis of the sequence shows that it contains the A and B consensus sequences found in many nucleotide-binding proteins. Twelve amino acids of ... More
Elimination of the hydroxyl groups in the ribose ring of ATP reduces its ability to phosphorylate the sarcoplasmic reticulum Ca(2+)-ATPase.
AuthorsCoan C, Amaral Júnior JA, Verjovski-Almeida S
JournalJ Biol Chem
PubMed ID8463222
'2''-Deoxyadenosine 5''-triphosphate, 3''-deoxyadenosine 5''-triphosphate, and 3''-amino-3''-deoxyadenosine 5''-triphosphate were substituted for ATP in the Ca2+ pumping cycle of the sarcoplasmic reticulum Ca(2+)-ATPase. The rate of phosphorylation of the enzyme decreased by more than an order of magnitude when either of the hydroxyl groups was eliminated from the ribose ring. This resulted ... More
Structural organization of chloroplast coupling factor.
AuthorsSnyder B, Hammes GG
JournalBiochemistry
PubMed ID2859887
'Fluorescence resonance energy transfer measurements have been used to construct spatial maps for the accessible sulfhydryl of the gamma subunit (dark site) and the essential tyrosine residue of the beta subunits relative to previously mapped sites on the H+-ATPase from chloroplasts. The extent of energy transfer was measured between a ... More
Interactions of nucleotide cofactors with the Escherichia coli replication factor DnaC protein.
AuthorsGalletto R, Rajendran S, Bujalowski W
JournalBiochemistry
PubMed ID11041861
'Quantitative analyses of the interactions of nucleotide cofactors with the Escherichia coli replicative factor DnaC protein have been performed using thermodynamically rigorous fluorescence titration techniques. This approach allowed us to obtain stoichiometries of the formed complexes and interaction parameters, without any assumptions about the relationship between the observed signal and ... More
Mitochondrial ATP synthase. Interaction of a synthetic 50-amino acid, beta-subunit peptide with ATP.
'A 50-amino acid peptide predicted by chemical modification studies of F1 and by comparison with adenylate kinase to comprise part of an ATP-binding domain within the beta-subunit of mitochondrial ATP synthase has been synthesized and purified. In the numbering system used for bovine heart beta, the peptide consists of amino ... More
Conformational changes in the unique loops bordering the ATP binding cleft of skeletal muscle myosin mediate energy transduction.
AuthorsMaruta S, Homma K
JournalJ Biochem (Tokyo)
PubMed ID11011153
'Myosin has three highly-conserved, unique loops [B (320-327), M (677-689), and N (127-136)] at the entrance of the ATP binding cleft, and we previously showed that the effects of actin are mediated by a conformational change in loop M [Maruta and Homma (1998) J. Biochem. 124, 528-533]. In the present ... More
Guinea-pig sympathetic neurons express varying proportions of two distinct P2X receptors.
AuthorsZhong Y, Dunn PM, Burnstock G
JournalJ Physiol
PubMed ID10699083
'1. Characterization of P2X receptors on neurons of guinea-pig superior cervical ganglion (SCG) has been carried out using a whole-cell voltage-clamp technique. 2. Application of ATP and alpha,beta-methylene ATP (alphabeta-MeATP) produced fast activating inward currents, which desensitized slowly. The maximum response to alphabeta-MeATP was 36 +/- 23 % (range 0.1-100 ... More
Excitation energy transfer studies on the proximity between SH1 and the adenosinetriphosphatase site in myosin subfragment 1.
AuthorsTao T, Lamkin M
JournalBiochemistry
PubMed ID6457630
'Excitation energy transfer studies were carried out to determine the distance between the adenosinetriphosphatase (ATPase) site and a unique "fast-reacting" sulfhydryl (referred to as SH1) in myosin subfragment 1. The fluorescent moiety of the probe N-(iodoacetyl)-N''-(5-sulfo-1-naphthyl)ethylene-diamine was used as the donor attached at SH1. The chromophoric nucleotide analogue 2''(3'')-0-(2,4,6-trinitrophenyl)adenosine 5''-diphosphate ... More
Effects of photo-oxidizing analogs of fluorescein on the sarcoplasmic reticulum Ca2+-ATPase. Functional consequences for substrate hydrolysis and effects on the partial reactions of the hydrolytic cycle.
AuthorsMignaco JA, Barrabin H, Scofano HM
JournalJ Biol Chem
PubMed ID8702486
'Erythrosin B was used to photo-oxidize the sarcoplasmic reticulum Ca2+-ATPase. The ATPase activity is rapidly and irreversibly inhibited by photo-oxidation with erythrosin. This inhibition is protected by the presence of ATP during the photo-oxidation period. After photo-oxidation, the steady-state phosphorylation by ATP remains almost unchanged, whereas phosphorylation by inorganic phosphate ... More
Trinitrophenyl-ATP and -ADP bind to a single nucleotide site on isolated beta-subunit of Escherichia coli F1-ATPase. In vitro assembly of F1-subunits requires occupancy of the nucleotide-binding site on beta-subunit by nucleoside triphosphate.
AuthorsRao R, Al-Shawi MK, Senior AE
JournalJ Biol Chem
PubMed ID2895769
'The stoichiometry of nucleotide binding to the isolated alpha- and beta-subunits of Escherichia coli F1-ATPase was investigated using two experimental techniques: (a) titration with fluorescent trinitrophenyl (TNP) derivatives of AMP, ADP, and ATP and (b) the centrifuge column procedure using the particular conditions of Khananshvili and Gromet-Elhanan (Khananshvili, D., and ... More
Thermophilic F1-ATPase is activated without dissociation of an endogenous inhibitor, epsilon subunit.
AuthorsKato Y, Matsui T, Tanaka N, Muneyuki E, Hisabori T, Yoshida M
JournalJ Biol Chem
PubMed ID9312092
'Subunit complexes (alpha3beta3gamma, alpha3beta3gammadelta, alpha3beta3gammaepsilon, and alpha3beta3gammadeltaepsilon) of thermophilic F1-ATPase were prepared, and their catalytic properties were compared to know the role of delta and epsilon subunits in catalysis. The presence of delta subunit in the complexes had slight inhibitory effect on the ATPase activity. The effect of epsilon subunit ... More
Catalytic cooperativity of beef heart mitochondrial F1-ATPase revealed by using 2',3'-O-(2,4,6-trinitrophenyl)-ATP as a substrate; an indication of mutually activating catalytic sites.
AuthorsMuneyuki E, Hisabori T, Allison WS, Jault JM, Sasayama T, Yoshida M
JournalBiochim Biophys Acta
PubMed ID7947899
'The interaction of 2'',3''-O-(2,4,6-trinitrophenyl)ATP (TNP-ATP) with bovine mitochondrial F1-ATPase (MF1) was examined under substoichiometric and stoichiometric conditions to investigate the relationship between the amount of bound TNP-AT(D)P and extent of inhibition on steady state ATP hydrolysis. The hydrolysis of bound TNP-ATP under substoichiometric condition proceeded slowly, with a first order ... More
Pyridoxal kinase. Structure and function.
AuthorsChurchich JE, Kim YT
JournalAnn N Y Acad Sci
PubMed ID2162645
'Chymotryptic digestion of sheep brain pyridoxal kinase, a dimer of identical subunits each of 40 kDa, yields two fragments of 24 and 16 kDa with concomitant loss of catalytic activity. These fragments were separated by HPLC and used for binding studies with ATP and pyridoxal analogues. The spectroscopic properties of ... More
Assessment of the number of nucleotide binding sites on chloroplast coupling factor 1 by the continuous variation method.
AuthorsMusier KM, Hammes GG
JournalBiochemistry
PubMed ID2904277
'The method of continuous variation (Job plot analysis) and difference absorbance spectroscopy were used to investigate the binding of 2''(3'')-(trinitrophenyl)-ADP and -ATP to chloroplast coupling factor 1 (CF1). Experiments performed at a low total concentration (30 microM) of nucleotide and enzyme binding sites (assuming three or four binding sites per ... More
Inhibition study of ADP,ATP transport in mitochondria with trinitrophenyl-modified substrates.
AuthorsSchlimme E, Boos KS, Onur G, Ponse G
JournalFEBS Lett
PubMed ID6301884
'The ADP,ATP carrier of rat liver mitochondria is specifically inhibited by Meisenheimer-type trinitrophenyl (TNP) derivatives of ADP and ATP. Due to a systematic inhibition study we could show that the TNP-moiety itself, even in the 2'', 3''-O-cyclic Meisenheimer complex, revealed no inhibition of mitochondrial ADP,ATP transport. Nucleosidic TNP-compounds are weak ... More
Absorbance and fluorescence properties of 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate bound to coupled and uncoupled Ca2+-ATPase of skeletal muscle sarcoplasmic reticulum.
AuthorsBerman MC
JournalJ Biol Chem
PubMed ID2946686
'Preincubation of skeletal muscle sarcoplasmic reticulum vesicles in the presence of the calcium chelator, [ethylenebis(oxyethylenenitrilo)tetraacetic acid] (EGTA), irreversibly uncouples calcium transport from ATP hydrolysis. Uncoupling cannot be explained by increased membrane permeability, but is associated with decreased capacity of the Ca2+-ATPase to bind noncatalytic, tightly bound ATP and ADP (Berman, ... More
Dehydroepiandrosterone potentiates native ionotropic ATP receptors containing the P2X2 subunit in rat sensory neurones.
AuthorsDe Roo M, Rodeau JL, Schlichter R
JournalJ Physiol
PubMed ID12844512
'We have studied the modulatory effect of dehydroepiandrosterone (DHEA), the most abundant neurosteroid produced by glial cells and neurones, on membrane currents induced by the activation of ionotropic ATP (P2X) receptors in neonatal rat dorsal root ganglion neurones. ATP (1 microM) induced three types of currents/responses termed F (fast and ... More
2' (or 3')-O-(2, 4, 6-trinitrophenyl)adenosine 5'-triphosphate as a probe for the binding site of heavy meromyosin ATPase.
AuthorsHiratsuka T
JournalJ Biochem (Tokyo)
PubMed ID131793
'1. From NMR, IR and visible absorption studies of 2''(or 3'')-O-(2, 4, 6-trinitrophenyl)-adenosine 5''-triphosphate (TNP-ATP), 2''(or 3'')-O-(2, 4, 6-trinitrophenyl) adenosine (TNP-Ad(, and 1-(2''-hydroxyethoxy)-2, 4, 6-trinitrobenzene (TNP-EG), it was concluded that there is an intramolecular interaction between the base and 2, 4, 6-trinitrophenyl (TNP) moieties in the TNP-ATP molecule. 2. A ... More
TNP-ATP-resistant P2X ionic current on the central terminals and somata of rat primary sensory neurons.
AuthorsTsuzuki K, Ase A, Séguéla P, Nakatsuka T, Wang CY, She JX, Gu JG
JournalJ Neurophysiol
PubMed ID12783957
'P2X receptors have been suggested to be expressed on the central terminals of A delta-afferent fibers innervating dorsal horn lamina V and play a role in modulating sensory synaptic transmission. These P2X receptors have been widely thought to be P2X2+3 receptors. However, we have recently found that P2X receptor-mediated modulation ... More
Analysis of the properties of the N-terminal nucleotide-binding domain of human P-glycoprotein.
AuthorsBooth CL, Pulaski L, Gottesman MM, Pastan I
JournalBiochemistry
PubMed ID10820025
'Human P-glycoprotein, the MDR1 gene product, requires both Mg(2+)-ATP binding and hydrolysis to function as a drug transporter; however, the mechanism(s) defining these events is not understood. In the present study, we explored the nature of Mg(2+)-ATP binding in the N-terminal nucleotide-binding domain of human P-glycoprotein and identified the minimal ... More
Fluorescence studies of threonine-promoted conformational transitions in aspartokinase I using the substrate analogue 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate.
AuthorsBroglie KE, Takahashi M
JournalJ Biol Chem
PubMed ID6313682
'The trinitrophenyl derivative of ATP, 2''(3'')-O-(2,4,6-trinitrophenyl) adenosine 5''-triphosphate, has been used as a spectroscopic probe to investigate threonine-promoted conformational changes in the aspartokinase region of aspartokinase-homoserine dehydrogenase I in an attempt to relate the structural effects of threonine binding to inhibition of enzymatic activity. Binding of this analogue substrate to ... More
ADP regulates movements of mitochondria in neurons.
AuthorsMironov SL
JournalBiophys J
PubMed ID17277190
'Mitochondria often reside in subcellular regions with high metabolic demands. We examined the mechanisms that can govern the relocation of mitochondria to these sites in respiratory neurons. Mitochondria were visualized using tetramethylrhodamineethylester, and their movements were analyzed by applying single-particle tracking. Intracellular ATP ([ATP](i)) was assessed by imaging the luminescence ... More
Stoichiometry and mechanism of assembly of SV40 T antigen complexes with the viral origin of DNA replication and DNA polymerase alpha-primase.
AuthorsHuang SG, Weisshart K, Gilbert I, Fanning E
JournalBiochemistry
PubMed ID9799495
'The interactions of simian virus 40 (SV40) large T antigen with DNA carrying the viral origin of DNA replication, as well as its interactions with cellular replication proteins, have been investigated by using fluorescent ATP analogues as specific probes. The enhanced fluorescence of 3''(2'')-O-(2,4, 6-trinitrophenyl)adenosine diphosphate (TNP-ADP) induced by T ... More
Calcium transport by sarcoplasmic reticulum Ca(2+)-ATPase. Role of the A domain and its C-terminal link with the transmembrane region.
AuthorsMöller JV, Lenoir G, Marchand C, Montigny C, le Maire M, Toyoshima C, Juul BS, Champeil P
JournalJ Biol Chem
PubMed ID12138099
'After treatment of sarcoplasmic reticulum Ca(2+)-ATPase with proteinase K (PK) in the presence of Ca(2+) and a protecting non-phosphorylated ligand (e.g. adenosine 5''-(beta,gamma-methylenetriphosphate), we were able to prepare in high yield an ATPase species that only differs from intact ATPase because of excision of the MAATE(243) sequence from the loop ... More
The use of 2',3'-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate for studies of nucleotide interaction with sarcoplasmic reticulum vesicles.
AuthorsWatanabe T, Inesi G
JournalJ Biol Chem
PubMed ID6214553
'The ATP analogue 2'',3''-O-(2,4,6-trinitrophenyl) adenosine 5''-triphosphate (TNP-ATP) was used to study nucleotide site stoichiometry and interactions in sarcoplasmic reticulum (SR) vesicles. The TNP absorption spectrum in the visible region undergoes a specific change upon binding of the nucleotide to SR ATPase. Equilibrium binding was therefore measured by differential spectrophotometry. In ... More
Interaction of 4-azido-2-nitrophenyl phosphate, a pseudosubstrate, with the sarcoplasmic reticulum Ca-ATPase.
AuthorsLacapère JJ, Garin J
JournalBiochemistry
PubMed ID8117720
'In the dark, 4-azido-2-nitrophenyl phosphate (ANPP) is a phosphate analog which behaves like a simple energy-rich phosphate donor for the sarcoplasmic reticulum (SR) Ca-ATPase. Like p-nitrophenyl phosphate (pNPP), ANPP is hydrolyzed by the enzyme only in the presence of calcium and magnesium (K0.5 and Vmax are 0.3 mM and 60 ... More
Characterization of ATP and DNA binding activities of TrwB, the coupling protein essential in plasmid R388 conjugation.
AuthorsMoncalián G, Cabezón E, Alkorta I, Valle M, Moro F, Valpuesta JM, Goñi FM, de La Cruz F
JournalJ Biol Chem
PubMed ID10593894
'TrwB is the conjugative coupling protein of plasmid R388. TrwBDeltaN70 contains the soluble domain of TrwB. It was constructed by deletion of trwB sequences containing TrwB N-proximal transmembrane segments. Purified TrwBDeltaN70 protein bound tightly the fluorescent ATP analogue TNP-ATP (K(s) = 8.7 microM) but did not show measurable ATPase or ... More
Fluoride binding to the calcium ATPase of sarcoplasmic reticulum converts its transport sites to a low affinity, lumen-facing form.
AuthorsMurphy AJ, Coll RJ
JournalJ Biol Chem
PubMed ID1387398
'The sarcoplasmic reticulum CaATPase forms an inactive complex with fluoride (CaATPase-F), which in the absence of calcium reactivates very slowly (t1/2 approximately 40 h at 25 degrees C). Reactivation is greatly accelerated (greater than 10(3)) by calcium in the millimolar range provided it has access to luminal sites of the ... More
Studies of the interactions of 2',3'-O-(2,4,6-trinitrocyclohexyldienylidine)adenosine nucleotides with the sarcoplasmic reticulum (Ca2+ + Mg2+)-ATPase active site.
AuthorsNakamoto RK, Inesi G
JournalJ Biol Chem
PubMed ID6142049
'The fluorescence of TNP-nucleotides bound to sarcoplasmic reticulum ATPase is enhanced upon formation of phosphorylated enzyme intermediate either with ATP in the presence of Ca2+ or, to a greater extent, with Pi in the absence of Ca2+. Binding of the TNP-nucleotides does not occur if the ATPase is labeled at ... More
Estimation of the distance change between cysteine-457 and the nucleotide binding site when sodium pump changes conformation from E1 to E2 by fluorescence energy transfer measurements.
AuthorsLin SH, Faller LD
JournalBiochemistry
PubMed ID8679600
'The first indication of the size of a conformational change implicated in ion transport by sodium pump has been obtained by measuring the change in efficiency of fluorescence energy transfer between two specific locations on the alpha-subunit. The donor (5''-(iodoacetamido)fluorescein) attaches covalently to cysteine-457, and the acceptor (2''(or 3'')-O-(trinitrophenyl)adenosine 5''-triphosphate) ... More
Insight into the uncoupling mechanism of sarcoplasmic reticulum ATPase using the phosphorylating substrate UTP.
AuthorsFortea MI, Soler F, Fernandez-Belda F
JournalJ Biol Chem
PubMed ID10777540
'Ca(2+) transport and UTP hydrolysis catalyzed by sarcoplasmic reticulum Ca(2+)-ATPase from skeletal muscle was studied. A passive Ca(2+) load inside microsomal vesicles clearly decreased the net uptake rate and the final accumulation of Ca(2+) but not the UTP hydrolysis rate, causing energy uncoupling. In the absence of passive leak, the ... More
Wheat germ translation initiation factor eIF4B affects eIF4A and eIFiso4F helicase activity by increasing the ATP binding affinity of eIF4A.
AuthorsBi X, Ren J, Goss DJ
JournalBiochemistry
PubMed ID10801326
'It has been proposed that, during translational initiation, structures in the 5'' untranslated region of mRNA are unwound. eIF4A, a member of the DEAD box family of proteins (those that contain a DEAD amino acid sequence), separately or in conjunction with other eukaryotic initiation factors, utilizes the energy from ATP ... More
Different biochemical modes of action of two irreversible H+,K(+)-ATPase inhibitors, omeprazole and E3810.
AuthorsMorii M, Takeguchi N
JournalJ Biol Chem
PubMed ID8408006
'Omeprazole and E3810 were found to inhibit gastric H+,K(+)-ATPase following different biochemical mechanisms. Effects of the specific binding of the inhibitors on the conformational state of the enzyme were studied by measuring the fluorescence of the enzyme labeled with fluorescein 5''-isothiocyanate. The absolute fluorescence level of the omeprazole-bound enzyme was ... More
Eukaryotic initiation factor 4A is the component that interacts with ATP in protein chain initiation.
AuthorsSeal SN, Schmidt A, Marcus A
JournalProc Natl Acad Sci U S A
PubMed ID6579543
'Protein synthesis in a resolved homogenate of wheat germ requires ATP and eight factors functioning at the level of protein chain initiation. To identify the component(s) interacting with ATP, the different factors were treated with the ATP affinity analogue 5''-p-fluorosulfonylbenzoyladenosine (FSBA) and tested for their function in protein synthesis. The ... More
Expression and function of P2X purinoceptors in rat histaminergic neurons.
AuthorsVorobjev VS, Sharonova IN, Haas HL, Sergeeva OA
JournalBr J Pharmacol
PubMed ID12642404
'(1) The pharmacology of ATP responses and the expression pattern of seven known subunits of the P2X receptor were investigated in individual histaminergic neurons of the tuberomamillary nucleus (TM). (2) ATP (3-1000 micro M) evoked fast non-desensitizing inward currents in TM neurons. 2-methylthioATP (2MeSATP) displayed the same efficacy but a ... More
Mitochondrial ATP synthase. Overexpression in Escherichia coli of a rat liver beta subunit peptide and its interaction with adenine nucleotides.
AuthorsGarboczi DN, Hullihen JH, Pedersen PL
JournalJ Biol Chem
PubMed ID2902092
'The C-terminal two-thirds of the rat liver ATP synthase beta subunit has been overexpressed and exported to the Escherichia coli periplasm under the direction of the alkaline phosphatase (phoA) promoter and leader peptide. The processed soluble protein contains the 358 amino acids from glutamate 122 to the rat liver beta ... More
Human biliverdin reductase is autophosphorylated, and phosphorylation is required for bilirubin formation.
AuthorsSalim M, Brown-Kipphut BA, Maines MD
JournalJ Biol Chem
PubMed ID11278740
'Biliverdin reductase (BVR) reduces heme oxygenase (HO) activity product, biliverdin, to bilirubin. BVR is unique in having dual pH/dual cofactor requirements. Using Escherichia coli-expressed human BVR and COS cells, we show that BVR is autophosphorylated and that phosphorylation is required for its activity. An "in blot" autophosphorylation assay showed that ... More
Overexpression in Escherichia coli and purification of an ATP-binding peptide from the yeast plasma membrane H(+)-ATPase.
AuthorsCapieaux E, Rapin C, Thinès D, Dupont Y, Goffeau A
JournalJ Biol Chem
PubMed ID8408044
'The two major hydrophilic domains from the Saccharomyces cerevisiae plasma membrane H(+)-ATPase fused to glutathione S-transferase have been expressed in Escherichia coli. The GST-L peptide contained the hydrophilic region from Ala340 to Ser660. The GST-SL peptide contained in addition the hydrophilic region Glu162 to Val276. After solubilization of the inclusion ... More
Nucleotide and Mg2+ dependency of the thermal denaturation of mitochondrial F1-ATPase.
AuthorsVillaverde J, Cladera J, Hartog A, Berden J, Padrós E, Duñach M
JournalBiophys J
PubMed ID9746539
'The influence of adenine nucleotides and Mg2+ on the thermal denaturation of mitochondrial F1-ATPase (MF1) was analyzed. Differential scanning calorimetry in combination with ATPase activity experiments revealed the thermal unfolding of MF1 as an irreversible and kinetically controlled process. Three significant elements were analyzed during the thermal denaturation process: the ... More
The ATPase activity of the ChlI subunit of magnesium chelatase and formation of a heptameric AAA+ ring.
AuthorsReid JD, Siebert CA, Bullough PA, Hunter CN
JournalBiochemistry
PubMed ID12779346
'The AAA(+) ATPase component of magnesium chelatase (ChlI) drives the insertion of Mg(2+) into protoporphyrin IX; this is the first step in chlorophyll biosynthesis. We describe the ATPase activity, nucleotide binding kinetics, and structural organization of the ChlI protein. A consistent reaction scheme arises from our detailed steady state description ... More
Expression of functional purinergic receptors in pulmonary neuroepithelial bodies and their role in hypoxia chemotransmission.
AuthorsFu XW, Nurse CA, Cutz E
JournalBiol Chem
PubMed ID15134341
'Adenine nucleotides act through specific cell surface receptors to invoke a variety of biological responses. Here we show that cells of neuroepithelial bodies (NEB), presumed O2 airway sensors in neonatal hamster lung, express functional P2X receptors (P2X-R). Positive immunostaining was detected in NEB cells using double-label immunohistochemistry with antibodies against ... More
Fluorescence studies on the nucleotide binding domains of the P-glycoprotein multidrug transporter.
AuthorsLiu R, Sharom FJ
JournalBiochemistry
PubMed ID9062112
'One of the major causes of multidrug resistance in human cancers is expression of the P-glycoprotein multidrug transporter, which acts as an efflux pump for a diverse range of natural products, chemotherapeutic drugs, and hydrophobic peptides. In the present study, fluorescence techniques were used to probe the nucleotide binding domains ... More
Reaction sequences for (Na+ + K+)-dependent ATPase hydrolytic activities: new quantitative kinetic models.
AuthorsRobinson JD, Leach CA, Davis RL, Robinson LJ
JournalBiochim Biophys Acta
PubMed ID3015217
'To delineate better the reaction sequence of the (Na+ + K+)-ATPase and illuminate properties of the active site, kinetic data were fitted to specific quantitative models. For the (Na+ + K+)-ATPase reaction, double-reciprocal plots of velocity against ATP (in the millimolar range), with a series of fixed KCl concentrations, are ... More
P2 purinoceptor-mediated control of rat cerebral (pial) microvasculature; contribution of P2X and P2Y receptors.
AuthorsLewis CJ, Ennion SJ, Evans RJ
JournalJ Physiol
PubMed ID10970432
'Purine and pyrimidine nucleotides evoke changes in the vascular tone of medium to large cerebral vessels through the activation of P2 purinoceptors. We have applied P2 receptor drugs to rat pial arterioles and measured changes in arteriole diameter (o.d. 40-84 micrometer at rest), and recorded currents from arteriolar smooth muscle ... More
Characterization of the tryptophan fluorescence from sarcoplasmic reticulum adenosinetriphosphatase by frequency-domain fluorescence spectroscopy.
AuthorsGryczynski I, Wiczk W, Inesi G, Squier T, Lakowicz JR
JournalBiochemistry
PubMed ID2525924
'We examined the tryptophan decay kinetics of sarcoplasmic reticulum Ca2+-ATPase using frequency-domain fluorescence. Consistent with earlier reports on steady-state fluorescence intensity, our intensity decays reveal a reproducible and statistically significant 2% increase in the mean decay time due to calcium binding to specific sites involved in enzyme activation. This Ca2+ ... More
Binding of the fluorescent substrate analogue 2',3'-O-(2,4,6-trinitrophenylcyclohexadienylidene)adenosine 5'-triphosphate to the gastric H+,K(+)-ATPase: evidence for cofactor-induced conformational changes in the enzyme.
AuthorsFaller LD
JournalBiochemistry
PubMed ID2159328
'TNP-ATP binds to the gastric H,K-ATPase with a 4.6-fold increase in fluorescence intensity and 10-nm blue shift that indicate a relatively hydrophobic protein environment. The fluorescence enhancement saturates and is compatible with binding to a single class of specific nucleotide sites with Kd less than 25 nM and N = ... More
Resonance energy transfer: methods and applications.
AuthorsWu P, Brand L
JournalAnal Biochem
PubMed ID8053542
'Resonance energy transfer is widely used in studies of biomolecular structure and dynamics. It provides information about distances on the order of 10 to 100 A and is thus suitable for investigating spatial relationships of interest in biochemistry. The information available from energy transfer studies has been enhanced by the ... More
Fluorescence resonance energy transfer between the nucleotide binding site and Cys-10 in G-actin and F-actin.
AuthorsMiki M, Barden JA, dos Remedios CG
JournalBiochim Biophys Acta
PubMed ID3089284
'Intramonomer fluorescence resonance energy transfer between the donor epsilon-ATP bound to the nucleotide site and the acceptor N-(4-dimethylamino-3,5-dinitrophenyl)maleimide (DDPM) or 4-dimethylaminophenyl-azophenyl-4''-maleimide bound to Cys-10 in G-actin was measured. The donor-acceptor distance was calculated to be about 40 A. The intermonomer energy transfer in F-actin occurring between epsilon-ADP and DABMI was ... More
Binding of 2'(3')-O-(2,4,6-trinitrophenyl)-adenosine 5'-diphosphate opens the pathway for protons through the chloroplast ATPase complex.
AuthorsWagner R, Ponse G, Strotmann H
JournalEur J Biochem
PubMed ID3023082
'The effect of 2''(3'')-O-(2,4,6-trinitrophenyl)-adenosine 5''-diphosphate (TNP-ADP) on photophosphorylation and on the proton conductivity of the thylakoid membrane has been investigated. The results show that TNP-ADP is a potent competitive inhibitor of photophosphorylation (Ki = 1-2 microM). Moreover, in the absence of ADP and Pi, TNP-ADP accelerates basal electron transport of ... More
Substrate and DNA binding to a 50-residue peptide fragment of DNA polymerase I. Comparison with the enzyme.
AuthorsMullen GP, Shenbagamurthi P, Mildvan AS
JournalJ Biol Chem
PubMed ID2684960
'The fluorescent nucleotide 2',3'-trinitrophenyl-ATP (TNP-ATP) binds at the triphosphate substrate binding site of the large (Klenow) fragment of DNA polymerase I (Pol I) as detected by direct binding studies measuring the increase in fluorescence of this ligand (n = 1.0, KD = 0.07 microM). The enzyme-TNP-ATP complex binds Mg2+ and ... More
Fluorescence resonance energy transfer between points on tropomyosin and actin in skeletal muscle thin filaments: does tropomyosin move?
AuthorsMiki M, Miura T, Sano K, Kimura H, Kondo H, Ishida H, Maéda Y
JournalJ Biochem (Tokyo)
PubMed ID9603999
'Fluorescence resonance energy transfer (FRET) spectroscopy has been used to determine spatial relationships between residues on tropomyosin and actin in reconstituted muscle thin filament, and to detect a positional change of tropomyosin relative to actin on the thin filament in the presence and absence of Ca2+ ions. In addition to ... More
Distinct roles of P2X receptors in modulating glutamate release at different primary sensory synapses in rat spinal cord.
'Using spinal cord slice preparations and patch-clamp recordings in lamina II and lamina V regions, we tested a hypothesis that P2X receptor subtypes differentially modulate glutamate release from primary afferent terminals innervating different sensory regions. We found that activation of P2X receptors by alpha,beta-methylene-ATP increased glutamate release onto >80% of ... More
Catalytic activity of the alpha3beta3gamma complex of F1-ATPase without noncatalytic nucleotide binding site.
AuthorsMatsui T, Muneyuki E, Honda M, Allison WS, Dou C, Yoshida M
JournalJ Biol Chem
PubMed ID9079639
'A mutant alpha3beta3gamma complex of F1-ATPase from thermophilic Bacillus PS3 was generated in which noncatalytic nucleotide binding sites lost their ability to bind nucleotides. It hydrolyzed ATP at an initial rate with cooperative kinetics (Km(1), 4 microM; Km(2), 135 microM) similar to the wild-type complex. However, the initial rate decayed ... More
Stoichiometry and mapping of the nucleotide sites in sarcoplasmic reticulum ATPase with the use of UTP.
AuthorsFerreira ST, Verjovski-Almeida S
JournalJ Biol Chem
PubMed ID2968343
'Purified sarcoplasmic reticulum ATPase was phosphorylated by either ATP or UTP under otherwise identical conditions. Calcium, pH, and nucleotide concentrations were adjusted to permit maximal steady-state accumulation of phosphoenzyme (EP). Either 4 or 8.5 nmol of EP/mg of protein were obtained with ATP or UTP, respectively. Tryptic digestion of phosphorylated ... More
Effects of nucleotides on the protein ligands to metals at the M2 and M3 metal-binding sites of the spinach chloroplast F1-ATPase.
AuthorsHouseman AL, LoBrutto R, Frasch WD
JournalBiochemistry
PubMed ID7880823
'We have identified the most probable protein ligands at the catalytic M3 and noncatalytic M2 metal-binding sites in the spinach chloroplast F1-ATPase (CF1) and here propose possible residues in the protein sequence for these ligands in latent CF1 in the absence of nucleotide. The changes in the metal ligands at ... More
ADP binding induces long-distance structural changes in the beta polypeptide of the chloroplast ATP synthase.
AuthorsMills DA, Seibold SA, Squier TC, Richter ML
JournalBiochemistry
PubMed ID7742314
'Binding of ADP to the beta polypeptide isolated from the catalytic F1 portion (CF1) of the chloroplast ATP synthase caused an increase of 10-20% in the steady state fluorescence intensity of fluorescent maleimides attached to the cysteine residue at position 63. Fluorescence lifetime distributions indicated that the beta polypeptide switched ... More
Excitation energy transfer studies of the proximity between tropomyosin and actin in reconstituted skeletal muscle thin filaments.
AuthorsTao T, Lamkin M, Lehrer SS
JournalBiochemistry
PubMed ID6224509
Limitations in linearized analyses of binding equilibria: binding of TNP-ATP to the H4-H5 loop of Na/K-ATPase.
AuthorsKubala M, Plásek J, Amler E
JournalEur Biophys J
PubMed ID12851794
'Binding of TNP-ATP [2'',3''- O-(2,4,6-trinitrophenyl)adenosine 5''-triphosphate, a fluorescent analogue of ATP] to the K605 protein was studied. This is an isolated N-domain in the cytoplasmic loop of the Na/K-ATPase alpha-subunit, lying between membrane-spanning segments 4 and 5 (sequence L(354)-I(604)). A titration equation is derived that explicitly makes it possible to ... More
The cystic fibrosis transmembrane conductance regulator. Effects of the most common cystic fibrosis-causing mutation on the secondary structure and stability of a synthetic peptide.
'Deletion of phenylalanine 508 (delta Phe-508) in the cystic fibrosis transmembrane conductance regulator (CFTR) protein causes approximately 70% of all cases of cystic fibrosis. This residue lies in a region of the protein that we have synthesized chemically and shown to bind adenine nucleotides (Thomas, P. J., Shenbagamurthi, P., Ysern, ... More
Single site hydrolysis of 2',3'-O-(2,4,6-trinitrophenyl)-ATP by the F1-ATPase from thermophilic bacterium PS3 is accelerated by the chase-addition of excess ATP.
AuthorsHisabori T, Muneyuki E, Odaka M, Yokoyama K, Mochizuki K, Yoshida M
JournalJ Biol Chem
PubMed ID1531655
'The interaction of 2'',3''-O-(2,4,6-trinitrophenyl)-adenosine 5''-triphosphate (TNP-ATP) and TNP-ADP to F1-ATPase from a thermophilic bacterium PS3 (TF1) was investigated. When TNP-ADP or TNP-ATP was added to the isolated alpha or beta subunit of TF1, characteristic difference spectra were generated for each subunit. Difference spectra generated on addition of these analogs to ... More
Structures of human thymidylate kinase in complex with prodrugs: implications for the structure-based design of novel compounds.
AuthorsOstermann N, Segura-Peña D, Meier C, Veit T, Monnerjahn C, Konrad M, Lavie A
JournalBiochemistry
PubMed ID12614151
'Nucleoside analogue prodrugs are dependent on efficient intracellular stepwise phosphorylation to their triphosphate form to become therapeutically active. In many cases it is this activation pathway that largely determines the efficacy of the drug. To gain further understanding of the determinants for efficient conversion by the enzyme thymidylate kinase (TMPK) ... More
The carboxyl termini of K(ATP) channels bind nucleotides.
AuthorsVanoye CG, MacGregor GG, Dong K, Tang L, Buschmann AS, Hall AE, Lu M, Giebisch G, Hebert SC
JournalJ Biol Chem
PubMed ID11956191
'ATP-sensitive potassium (K(ATP)) channels are expressed in many excitable, as well as epithelial, cells and couple metabolic changes to modulation of cell activity. ATP regulation of K(ATP) channel activity may involve direct binding of this nucleotide to the pore-forming inward rectifier (Kir) subunit despite the lack of known nucleotide-binding motifs. ... More
Reaction of a carbodiimide adduct of ATP at the active site of sarcoplasmic reticulum calcium ATPase.
AuthorsMurphy AJ
JournalBiochemistry
PubMed ID2148693
'An adduct of a carbodiimide and ATP was synthesized from 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC) and the nucleotide. Despite its limited stability (t1/2 for hydrolysis of about 5 min at 25 degrees C), it was shown to react with and inactivate the calcium ATPase of sarcoplasmic reticulum in its vesicular, nonionic detergent-solubilized and ... More
Both the ADP receptors P2Y1 and P2Y12, play a role in controlling shape change in human platelets.
AuthorsJagroop IA, Burnstock G, Mikhailidis DP
JournalPlatelets
PubMed ID12623443
'Two types of ADP receptors, P2Y(1) and P2Y(12) are involved in platelet aggregation. The P2X(1) receptor is also present but its role, in terms of platelet function, is not yet defined. The aim of this study was to establish if the ADP receptors, P2Y(1,) P2Y(12) and P2X(1) play a role ... More
Localization of site-specific probes on the Ca-ATPase of sarcoplasmic reticulum using fluorescence energy transfer.
AuthorsSquier TC, Bigelow DJ, Garcia de Ancos J, Inesi G
JournalJ Biol Chem
PubMed ID2951372
'Highly reactive sulfhydryls, previously labeled with an iodoacetamide spin label on the Ca-ATPase of sarcoplasmic reticulum, were labeled with the fluorescent probe, 5-(2-[iodoacetyl)amino)ethyl)aminonaphthalene-1-sulfonic acid (IAEDANS), without loss of enzymatic activity. We have selectively measured the apparent distance of the more reactive site, relative to other site-specific probes at both the ... More
Substrate binding-induced alteration of nucleotide binding site properties of chloroplast coupling factor 1.
AuthorsShapiro AB, McCarty RE
JournalJ Biol Chem
PubMed ID2137822
'Two adenine nucleotide binding sites of chloroplast coupling factor 1 (CF1) were shown previously to switch their properties after exposure of the enzyme to Mg2(+)-ATP or Ca2(+)-ATP (Shapiro, A. B., and McCarty, R. E. (1988) J. Biol. Chem. 263, 14160-14165). The change in binding properties was monitored by fluorescence resonance ... More
Probing the nucleotide binding domain of the osmoregulator EnvZ using fluorescent nucleotide derivatives.
AuthorsPlesniak L, Horiuchi Y, Sem D, Meinenger D, Stiles L, Shaffer J, Jennings PA, Adams JA
JournalBiochemistry
PubMed ID12437344
'EnvZ is a histidine protein kinase important for osmoregulation in bacteria. While structural data are available for this enzyme, the nucleotide binding pocket is not well characterized. The ATP binding domain (EnvZB) was expressed, and its ability to bind nucleotide derivatives was assessed using equilbrium and stopped-flow fluorescence spectroscopy. The ... More
Identification of active site residues in mevalonate diphosphate decarboxylase: implications for a family of phosphotransferases.
AuthorsKrepkiy D, Miziorko HM
JournalProtein Sci
PubMed ID15169949
'A combination of sequence homology analyses of mevalonate diphosphate decarboxylase (MDD) proteins and structural information for MDD leads to the hypothesis that Asp 302 and Lys 18 are active site residues in MDD. These residues were mutated to replace acidic/basic side chains and the mutant proteins were isolated and characterized. ... More
Fluorescence changes of a label attached near the myosin active site on nucleotide binding in rat skeletal muscle fibres.
AuthorsFujita S, Nawata T, Yamada K
JournalJ Physiol
PubMed ID10066911
'1. Trinitrophenyl AMP (TNP-AMP) in the concentration range 10-300 microM induced an increase in fluorescence intensity at around 530 nm in skinned skeletal muscle fibres freshly obtained from rat psoas muscle. 2. The fluorescence intensity of the fibres depended on TNP-AMP concentration up to approximately 200 microM. The Kd of ... More
Expression and characterization of the NBD1-R domain region of CFTR: evidence for subunit-subunit interactions.
AuthorsNeville DC, Rozanas CR, Tulk BM, Townsend RR, Verkman AS
JournalBiochemistry
PubMed ID9485388
'To study interactions between the contiguous NBD1 and R domains of CFTR, wild-type and DeltaF508 NBD1-R (amino acids 404-830, in fusion with His6 tag) were expressed as single proteins in Escherichia coli. NBD1-R (10-25 mg/L culture) was purified from inclusion bodies in 8 M urea by Ni-affinity chromatography, and renatured ... More
Characterization of 2'(3')-trinitrophenyl-ATP as an inhibitor of ATP-dependent initiation complex formation between the DNA polymerase III holoenzyme and primed DNA.
AuthorsOberfelder R, McHenry CS
JournalJ Biol Chem
PubMed ID3549726
'We have identified 2''(3'')-trinitrophenyl-ATP to be an inhibitor of the ATP-dependent initiation complex formation reaction between the Escherichia coli DNA polymerase III holoenzyme and primed DNA. The inhibitor is specific for the initiation stage; once initiation complexes are formed the subsequent elongation reaction is unaffected. Three ATP-dependent DNA polymerase III ... More
Localization of a polynucleotide binding region in the HIV-1 reverse transcriptase: implications for primer binding.
AuthorsSobol RW, Suhadolnik RJ, Kumar A, Lee BJ, Hatfield DL, Wilson SH
JournalBiochemistry
PubMed ID1718424
'Properties of primer recognition by purified human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) p66 homodimer have been investigated. Earlier studies had shown that RNA-directed DNA synthesis catalyzed by HIV-1 RT proceeds by an ordered mechanism in which template-primer combines with the free enzyme to form the first complex ... More
The dnaB protein of Escherichia coli: mechanism of nucleotide binding, hydrolysis, and modulation by dnaC protein.
AuthorsBiswas EE, Biswas SB, Bishop JE
JournalBiochemistry
PubMed ID3026453
'The mechanism of nucleotide binding and hydrolysis by dnaB protein and dnaB X dnaC protein complex has been studied by using fluorescent nucleotide analogues. Binding of trinitrophenyladenosine triphosphate (TNP-ATP) or the corresponding diphosphate (TNP-ADP) results in a blue shift of the emission maximum and a severalfold amplification of the fluorescence ... More
Light-dependent inactivation of (Na+ + K+)-ATPase with a new photoaffinity reagent, chromium arylazido-beta-alanyl ATP.
AuthorsMunson KB
JournalJ Biol Chem
PubMed ID6259155
Preparation and properties of 2'(or 3')-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate, an analog of adenosine triphosphate.
AuthorsHiratsuka T, Uchida K
JournalBiochim Biophys Acta
PubMed ID4270904
Heterogeneous reaction of heavy meromyosin ATPase with 2' (or 3')-O-(2,4,6-trinitrophenyl)adenosine 5'-5'-triphosphate.
AuthorsHiratsuka T, Uchida K, Yagi K
JournalJ Biochem (Tokyo)
PubMed ID144118
Titration of the nucleotide binding sites of sarcoplasmic reticulum Ca2+ -ATPase with 2',3'-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate and 5'-diphosphate.
AuthorsDupont Y, Chapron Y, Pougeois R
JournalBiochem Biophys Res Commun
PubMed ID6214259
Binding of TNP-ATP to IAF-labeled Na+/K(+)-ATPase as examined by fluorescence quenching.
AuthorsPratap PR, Hellen EH, Palit A
JournalAnn N Y Acad Sci
PubMed ID9405840
Fluorescence quenching of IAF-Na+/K(+)-ATPase via energy transfer to TNP-labeled nucleotide.
AuthorsHellen EH, Pratap PR
JournalAnn N Y Acad Sci
PubMed ID9405839
Structure of actin observed by fluorescence resonance energy transfer spectroscopy.
AuthorsMiki M, O'Donoghue SI, Dos Remedios CG
JournalJ Muscle Res Cell Motil
PubMed ID1534564
Myosin structure. Proximity measurements by fluorescence energy transfer.
AuthorsHaugland RP
JournalJ Supramol Struct
PubMed ID127888
Stoichiometric binding of 2'(or 3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate to bovine heart cytochrome c oxidase.
AuthorsReimann A, Kadenbach B
JournalFEBS Lett
PubMed ID1322834
The binding of 2'(or 3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP) to isolated bovine heart cytochrome c oxidase (COX) was studied by following its specific spectral change at 510 nm. The quantitative titration revealed two binding sites for TNP-ATP per monomer COX with a Kd of 1.6 microM. ... More
Synthesis of novel fluorescent-labelled dinucleoside polyphosphates.
AuthorsWright M, Miller AD
JournalBioorg Med Chem Lett
PubMed ID15125938
A novel tandem synthetic-biosynthetic procedure is described for the synthesis of four new fluorescent dinucleoside polyphosphates: mant-Ap4A, mant-AppCH2ppA, TNP-Ap4A and TNP-AppCH2ppA. These compounds are expected to supplement the existing etheno (epsilon) and 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) labelled derivatives, being the fluorescent probes of choice to investigate polyphosphate/enzyme binding behaviour. ... More
Inactivation of Ca2(+)-, Na+K(+)-, and H+K(+)-ATPases with a carbodiimide derivative of ATP.
AuthorsMurphy AJ
JournalFEBS Lett
PubMed ID2158904
The gamma-P adduct of ATP with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (ATP-EDC) was synthesized and incubated with the Ca-ATPase of sarcoplasmic reticulum with the result that time-dependent complete loss of the enzyme's activity occurred. The inactivation required calcium and magnesium while ATP had a protective effect. ATP-EDC incubation with the NaK-ATPase and HK-ATPase produced ... More
Characterisation of ATP binding inhibition to the sarcoplasmic reticulum Ca(2+)-ATPase by thapsigargin.
AuthorsDeJesus F, Girardet JL, Dupont Y
JournalFEBS Lett
PubMed ID8405462
The inhibition of Ca(2+)-ATPase of sarcoplasmic reticulum by thapsigargin has been reported to be associated with a suppression of calcium binding to the high affinity transport sites. We report here that thapsigargin also acts as an inhibitor of ATP binding by reducing its apparent affinity by about two orders of ... More
On the mechanism of energy transduction in myosin subfragment 1.
It is proposed that the myosin subfragment 1 moiety of the muscle contractile apparatus is a self-contained "engine" whose operational plan is based on the interactive nature of ATP (or degradation intermediate) binding and actin binding, made possible by an intersite communication system. It is suggested that the spatial information ... More
Fluorescence properties of 2' (or 3')-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate and its use in the study of binding to heavy meromyosin ATPase.
AuthorsHiratsuka T
JournalBiochim Biophys Acta
PubMed ID11824
2' (or 3')-O-(2,4,6-Trinitrophenyl) adenosine 5'-triphosphate (N3ph-ATP), which contains a Meisenheimer complex moiety, is one of the class of compounds which do not fluoresce in water but fluoresce both in low polarity solvents and when bound to the protein molecule. Fluorescence intensity of N3ph-ATP in the range of 540 nm, when ... More
Affinity of nucleotides for the active site of detergent-solubilized sarcoplasmic reticulum CaATPase.
AuthorsColl RJ, Murphy AJ
JournalBiochem Biophys Res Commun
PubMed ID2943280
A dye displacement method was developed for determination of the affinities of compounds toward the active site of the detergent-solubilized sarcoplasmic reticulum CaATPase. Titration of the enzyme with 2',3'-O-(2,4,6-trinitrophenyl)-ADP resulted in a large absorbance difference in the visible spectrum. Subsequent addition of compounds which bind to the active site causes ... More
Fluorescent and colored trinitrophenylated analogs of ATP and GTP.
AuthorsHiratsuka T
JournalEur J Biochem
PubMed ID12919312
Fluorescent and colored trinitrophenylated (TNP) analogs of ATP and GTP can interact with nucleotide-requiring enzymes and proteins as a substitute for the parent nucleotide. These analogs have strong binding affinities for most nucleotide-requiring systems. Their bindings are easily detected by absorption and fluorescence changes in the visible region. Recent years ... More
Structural mapping of cysteine-63 of the chloroplast ATP synthase beta subunit.
AuthorsColvert KK, Mills DA, Richter ML
JournalBiochemistry
PubMed ID1533153
The single sulfhydryl residue (cysteine-63) of the beta subunit of the chloroplast ATP synthase F1 (CF1) was accessible to labeling reagents only after removal of the beta subunit from the enzyme complex. This suggests that cysteine-63 may be located at an interface between the beta and the alpha subunits of ... More
Binding of 2'(3')-O-(2,4-6-trinitrophenyl) ADP to soluble alpha beta protomers of Na, K-ATPase modified with fluorescein isothiocyanate. Evidence for two distinct nucleotide sites.
AuthorsWard DG, Cavieres JD
JournalJ Biol Chem
PubMed ID8647832
The overall reaction of well-defined solubilized protomers of Na,K-ATPase (one alpha plus one beta subunit) retains the dual ATP dependence observed with the membrane-bound enzyme, with distinctive ATP effects in the submicromolar and submillimolar ranges (Ward, D. G., and Cavieres, J. D. (1993) Proc. Natl. Acad. Sci. U. S. A. ... More
ATP analogues at a glance.
AuthorsBagshaw C,
JournalJ Cell Sci
PubMed ID11171313
ATP has long been known to play a central role in the energetics of cells both in transduction mechanisms and in metabolic pathways, and is involved in regulation of enzyme, channel and receptor activities. Numerous ATP analogues have been synthesised to probe the role of ATP in biosystems (Yount, 1975; ... More
Trinitrophenyl-ATP binding to the ArsA protein: the catalytic subunit of an anion pump.
AuthorsKarkaria CE, Rosen BP
JournalArch Biochem Biophys
PubMed ID1832838
The ars operon of the conjugative R-factor R773 confers resistance to arsenicals by coding for an anion pump for extrusion of arsenicals from cells of Escherichia coli. Extrusion of arsenite requires only two polypeptides, the ArsA and ArsB proteins. Purified ArsA protein exhibits oxyanion-stimulated ATPase activity and has been shown ... More
The binding of the fluorescent ATP analogue 2'(3')-trinitrophenyladenosine-5'-triphosphate to rat liver fatty acid-binding protein.
AuthorsSheridan M, Wilton DC
JournalFEBS Lett
PubMed ID1468590
The less polar fluorescent analogue of ATP, 2'(3')-trinitrophenyl-5'-triphosphate bound to rat liver fatty acid-binding protein with high affinity (Kd 6.3 x 10(-6) M) and 1:1 molar stoichiometry. This probe bound to the fatty acid binding site of the protein and was displaced by oleic acid and oleoyl CoA. High concentrations ... More
Nucleotide binding by the histidine kinase CheA.
AuthorsBilwes AM, Quezada CM, Croal LR, Crane BR, Simon MI
JournalNat Struct Biol
PubMed ID11276258
To probe the structural basis for protein histidine kinase (PHK) catalytic activity and the prospects for PHK-specific inhibitor design, we report the crystal structures for the nucleotide binding domain of Thermotoga maritima CheA with ADP and three ATP analogs (ADPNP, ADPCP and TNP-ATP) bound with either Mg(2+) or Mn(2+). The ... More
Selective modification of an alpha subunit of chloroplast coupling factor 1.
AuthorsNalin CM, Snyder B, McCarty RE
JournalBiochemistry
PubMed ID2859886
Lucifer yellow (4-amino-N-[3-(vinylsulfonyl)phenyl]naphthalimide-3,6-disulf onate), a fluorescent probe that can react covalently with sulfhydryl or amino groups, has been used to modify chloroplast coupling factor 1 (CF1). Conditions are described under which Lucifer yellow selectively labels the alpha subunit of CF1 to the extent of about 1 mol of probe per ... More
Binding of TNP-ATP and TNP-ADP to the non-catalytic sites of Escherichia coli F1-ATPase.
AuthorsWeber J, Senior AE
JournalFEBS Lett
PubMed ID9257714
Using site-directed-tryptophan fluorescence, parameters for equilibrium binding of (Mg)TNP-ATP and (Mg)TNP-ADP to non-catalytic sites of Escherichia coli F1-ATPase were determined. All three non-catalytic sites showed the same affinity for MgTNP-ATP (Kd = 0.2 microM) or MgTNP-ADP (Kd = 6.5 microM) whereas even at concentrations of 100 microM no binding of ... More
Competitive binding of ATP and the fluorescent substrate analogue 2',3'-O-(2,4,6-trinitrophenylcyclohexadienylidine) adenosine 5'-triphosphate to the gastric H+,K+-ATPase: evidence for two classes of nucleotide sites.
AuthorsFaller LD
JournalBiochemistry
PubMed ID2551380
ATP and the fluorescent substrate analogue TNP-ATP bind competitively to the gastric H,-K-ATPase. Substrate and product completely reverse the fluorescence enhancement caused by TNP-ATP binding to the enzyme. The fluorophore is displaced monophasically from apoenzyme. However, ATP displaces TNP-ATP from the Mg2+-quenched state in two steps of equal amplitude. The ... More