Sliding distance per ATP molecule hydrolyzed by myosin heads during isotonic shortening of skinned muscle fibers.
AuthorsHiguchi H, Goldman YE
JournalBiophys J
PubMed ID8534820
'We measured isotonic sliding distance of single skinned fibers from rabbit psoas muscle when known and limited amounts of ATP were made available to the contractile apparatus. The fibers were immersed in paraffin oil at 20 degrees C, and laser pulse photolysis of caged ATP within the fiber initiated the ... More
Rotational dynamics of actin-bound intermediates in the myosin ATPase cycle.
AuthorsBerger CL, Thomas DD
JournalBiochemistry
PubMed ID1657157
'We have used saturation-transfer electron paramagnetic resonance (ST-EPR) to detect the microsecond rotational motions of spin-labeled myosin subfragment one (MSL-S1) bound to actin in the presence of the ATP analogues AMPPNP (5''-adenylylimido diphosphate) and ATP gamma S [adenosine 5''-O-(3-thiotriphosphate)], which are believed to trap myosin in strongly and weakly bound ... More
Sodium pump-mediated ATP:ADP exchange. The sided effects of sodium and potassium ions.
AuthorsKaplan JH
JournalJ Gen Physiol
PubMed ID6294224
'Resealed human red cell ghosts containing caged ATP (Kaplan et al., 1978) and [3H]ADP were irradiated at 340 nm. The photochemical release of free ATP initiated a rapid transphosphorylation reaction (ATP:ADP exchange), a component of which is inhibited by ouabain. The reaction rate was measured by following the rate of ... More
Na+ movement in a single turnover of the Na pump.
AuthorsForbush B
JournalProc Natl Acad Sci U S A
PubMed ID6089192
'Ouabain-sensitive 22Na efflux from right-side-out membrane vesicles prepared from dog kidney has been examined with a time resolution of 30 msec. The vesicles are preloaded with 22Na and caged ATP [P3-1-(2-nitro)phenylethyl adenosine triphosphate], so that transport by the Na pump can be initiated by light. After a brief illumination, which ... More
Electrical currents associated with nucleotide transport by the reconstituted mitochondrial ADP/ATP carrier.
AuthorsBrustovetsky N, Becker A, Klingenberg M, Bamberg E
JournalProc Natl Acad Sci U S A
PubMed ID8570612
'The electrophoretic export of ATP against the import of ADP in mitochondria bridges the intra- versus extramitochondrial ATP potential gap. Here we report that the electrical nature of the ADP/ATP exchange by the mitochondrial ADP/ATP carrier (AAC) can be directly studied by measuring the electrical currents via capacitive coupling of ... More
Adenosine triphosphate and the late steps in calcium-dependent exocytosis at a ribbon synapse.
AuthorsHeidelberger R
JournalJ Gen Physiol
PubMed ID9450941
'The ATP dependence of the kinetics of Ca2+-dependent exocytosis after flash photolysis of caged Ca2+ was studied by capacitance measurements with submillisecond resolution in single synaptic terminals of retinal bipolar neurons. After control experiments verified that this combination of techniques is valid for the study of exocytosis in synaptic terminals, ... More
Cryophotolysis of caged compounds: a technique for trapping intermediate states in protein crystals.
AuthorsUrsby T, Weik M, Fioravanti E, Delarue M, Goeldner M, Bourgeois D
JournalActa Crystallogr D Biol Crystallogr
PubMed ID11914484
'Caged compounds in combination with protein crystallography represent a valuable tool in studies of enzyme reaction intermediates. To date, photochemical triggering of reactions has been performed close to room temperature. Synchronous reaction initiation has only been achieved with enzymes of relatively slow turnover (<0.1 s(-1)) and caged compounds of high ... More
A birefringence study of changes in myosin orientation during relaxation of skinned muscle fibers induced by photolytic ATP release.
AuthorsPeckham M, Ferenczi MA, Irving M
JournalBiophys J
PubMed ID7811926
'The birefringence of isolated skinned fibers from rabbit psoas muscle was measured continuously during relaxation from rigor produced by photolysis of caged ATP at sarcomere length 2.8-2.9 microns, ionic strength 0.1 M, 15 degrees C. Birefringence, the difference in refractive index between light components polarized parallel and perpendicular to the ... More
The use of caged adenine nucleotides and caged phosphate in intact skeletal muscle fibres of the mouse.
AuthorsAllen DG, Lännergren J, Westerblad H
JournalActa Physiol Scand
PubMed ID10610612
'The effects of 1-(2-nitrophenyl)ethyl ester of ATP (NPE-caged ATP), NPE-caged ADP, NPE-caged phosphate (Pi) and desoxybenzoinyl phosphate (desyl-caged Pi) on mouse skeletal muscle function were studied. All these caged compounds, when microinjected into intact single mouse muscle fibres, reduced the myoplasmic calcium during a tetanus (tetanic [Ca2+]i) and reduced force. ... More
Kinetics of ATP hydrolysis and tension production in skinned cardiac muscle of the guinea pig.
AuthorsBarsotti RJ, Ferenczi MA
JournalJ Biol Chem
PubMed ID3182811
'The kinetics of ATP hydrolysis and tension responses were studied simultaneously in a permeabilized preparation of cardiac tissue of the guinea pig. This was achieved by combining laserflash photolysis of P3-1-(2-nitrophenyl)ethyladenosine 5''-triphosphate ("caged-ATP") and a rapid freezing technique. In the presence of calcium ions, tension increased following the photolytic production ... More
A pre-active attached state of myosin heads in rat skeletal muscles.
AuthorsYagi N, Horiuti K, Takemori S
JournalJ Muscle Res Cell Motil
PubMed ID9477379
'X-ray diffraction experiments were performed on rat soleus and psoas muscles using synchrotron radiation from a multipole wiggler. The X-ray diffraction patterns were recorded at a time resolution of 16.7 ms with an X-ray image intensifier and a CCD video camera. A bundle of 10-15 muscle fibres was activated by ... More
Initiation of active contraction by photogeneration of adenosine-5'-triphosphate in rabbit psoas muscle fibres.
AuthorsGoldman YE, Hibberd MG, Trentham DR
JournalJ Physiol
PubMed ID6481646
'Mechanical and biochemical descriptions of the muscle cross-bridge cycle have been correlated. Skinned muscle fibres of rabbit psoas muscle in rigor were incubated in solutions containing approximately equal to 30 microM-Ca2+ ions and P3-1-(2-nitro)phenylethyladenosine-5'-triphosphate, 'caged ATP', an inert photolabile precursor of ATP. ATP was liberated from caged ATP within the ... More
Rotational dynamics of spin-labeled F-actin during activation of myosin S1 ATPase using caged ATP.
AuthorsOstap EM, Thomas DD
JournalBiophys J
PubMed ID1651780
'The most probable source of force generation in muscle fibers in the rotation of the myosin head when bound to actin. This laboratory has demonstrated that ATP induces microsecond rotational motions of spin-labeled myosin heads bound to actin (Berger, C. L. E. C. Svensson, and D. D. Thomas. 1989. Proc. ... More
Conformational transitions and change translocation by the Na,K pump: comparison of optical and electrical transients elicited by ATP-concentration jumps.
AuthorsStürmer W, Apell HJ, Wuddel I, Läuger P
JournalJ Membr Biol
PubMed ID2552127
'The electrogenic properties of the Na,K-ATPase were studied by correlating transient electrical events in the pump molecule with conformational transitions elicited by an ATP-concentration jump. Flat membrane fragments containing a high density (approximately 8000 microm(-2)) of oriented Na,K-ATPase molecules were bound to a planar lipid bilayer acting as a capacitive ... More
Kinetics of contraction initiated by flash photolysis of caged adenosine triphosphate in tonic and phasic smooth muscles.
AuthorsHoriuti K, Somlyo AV, Goldman YE, Somlyo AP
JournalJ Gen Physiol
PubMed ID2559141
'Laser flash photolysis of caged adenosine triphosphate (ATP), in the presence of Ca2+, was used to examine the time course of isometric force development from rigor states in glycerinated tonic (rabbit trachealis) and phasic (guinea-pig ileum and portal vein) smooth muscles. Photolytic liberation of ATP from caged ATP initiated force ... More
Photolabile chelators for the rapid photorelease of divalent cations.
AuthorsKaplan JH, Ellis-Davies GC
JournalProc Natl Acad Sci U S A
PubMed ID3137570
'The properties of a recently synthesized photolabile chelator for divalent cations are described, the affinity of which for Ca2+ changes by some 5 orders of magnitude on illumination. The compound 1-(2-nitro-4,5-dimethoxyphenyl)-N,N,N'',N''-tetrakis[(oxycarbonyl)me thyl]-1,2-ethanediamine (DM-nitrophen) binds Ca2+ (Kd approximately 5.0 x 10(-9) M) and Mg2+ (Kd approximately 2.5 x 10(-6) M) with ... More
Current transients generated by the Na+/K+-ATPase after an ATP concentration jump: dependence on sodium and ATP concentration.
AuthorsBorlinghaus R, Apell HJ
JournalBiochim Biophys Acta
PubMed ID2451535
'Planar membrane fragments containing a high density of oriented Na+/K+-ATPase molecules are bound to planar lipid bilayers. ATP is released in the aqueous solution within milliseconds from an inactive, photolabile precursor (''caged ATP'') by an intense light flash. By this ATP-concentration jump a large number of pump molecules is activated ... More
Fast charge translocations associated with partial reactions of the Na,K-pump: I. Current and voltage transients after photochemical release of ATP.
AuthorsBorlinghaus R, Apell HJ, Läuger P
JournalJ Membr Biol
PubMed ID3041002
'Nonstationary electric currents are described which are generated by the Na,K-pump. Flat membrane sheets 0.2-1 micron in diameter containing a high density of oriented Na,K-ATPase molecules are bound to a planar lipid bilayer acting as a capacitive electrode. In the aqueous phase adjacent to the bound membrane sheets, ATP is ... More
Investigating structural changes induced by nucleotide binding to RecA using difference FTIR.
AuthorsButler BC, Hanchett RH, Rafailov H, MacDonald G
JournalBiophys J
PubMed ID11916875
'Nucleotide binding to RecA results in either the high-DNA affinity form (Adenosine 5''-triphosphate (ATP)-bound) or the more inactive protein conformation associated with a lower affinity for DNA (Adenosine 5''-diphosphate (ADP)-bound). Many of the key structural differences between the RecA-ATP and RecA-ADP bound forms have yet to be elucidated. We have ... More
The kinetics of magnesium adenosine triphosphate cleavage in skinned muscle fibres of the rabbit.
AuthorsFerenczi MA, Homsher E, Trentham DR
JournalJ Physiol
PubMed ID6611412
'The time course of magnesium adenosine triphosphate (Mg ATP) cleavage in chemically skinned muscle fibres of the rabbit was measured by a method in which Mg ATP cleavage was initiated by photolytic release of ATP from P3-1-(2-nitro)phenylethyladenosine 5'-triphosphate (caged ATP) and terminated by rapid freezing 50 ms to 8 s ... More
Time-resolved charge translocation by the Ca-ATPase from sarcoplasmic reticulum after an ATP concentration jump.
AuthorsHartung K, Froehlich JP, Fendler K
JournalBiophys J
PubMed ID9168027
'Time-resolved measurements of currents generated by Ca-ATPase from fragmented sarcoplasmic reticulum (SR) are described. SR vesicles spontaneously adsorb to a black lipid membrane acting as a capacitive electrode. Charge translocation by the enzyme is initiated by an ATP concentration jump performed by the light-induced conversion of an inactive precursor (caged ... More
The myosin cross-bridge cycle and its control by twitchin phosphorylation in catch muscle.
AuthorsButler TM, Narayan SR, Mooers SU, Hartshorne DJ, Siegman MJ
JournalBiophys J
PubMed ID11159412
'The anterior byssus retractor muscle of Mytilus edulis was used to characterize the myosin cross-bridge during catch, a state of tonic force maintenance with a very low rate of energy utilization. Addition of MgATP to permeabilized muscles in high force rigor at pCa > 8 results in a rapid loss ... More
Replacement of glycine 232 by aspartic acid in the KdpA subunit broadens the ion specificity of the K(+)-translocating KdpFABC complex.
AuthorsSchrader M, Fendler K, Bamberg E, Gassel M, Epstein W, Altendorf K, Dröse S
JournalBiophys J
PubMed ID10920013
'Replacement of glycine residue 232 with aspartate in the KdpA subunit of the K(+)-translocating KdpFABC complex of Escherichia coli leads to a transport complex that has reduced affinity for K(+) and has lost the ability to discriminate Rb(+) ions (, J. Biol. Chem. 270:6678-6685). This glycine residue is the first ... More
Activation of the Drosophila TRP and TRPL channels requires both Ca2+ and protein dephosphorylation.
AuthorsAgam K, Frechter S, Minke B
JournalCell Calcium
PubMed ID14706283
'The Transient Receptor Potential (TRP) proteins constitute a large and diverse family of channel proteins, which is conserved through evolution. TRP channel proteins have critical functions in many tissues and cell types, but their gating mechanism is an enigma. In the present study patch-clamp whole-cell recordings was applied to measure ... More
Rotational dynamics of actin-bound myosin heads in active myofibrils.
AuthorsBerger CL, Thomas DD
JournalBiochemistry
PubMed ID8385491
'We have used saturation-transfer electron paramagnetic resonance (ST-EPR) to measure the submillisecond rotational motions of actin-bound myosin heads in active myofibrils. The cross-bridges were spin-labeled with a maleimide nitroxide derivative (MSL) that has previously been shown to undergo microsecond rotational motions on actin-bound myosin heads in solution during steady-state ATPase ... More
Microtubule-motor activity of a yeast centromere-binding protein complex.
AuthorsHyman AA, Middleton K, Centola M, Mitchison TJ, Carbon J
JournalNature
PubMed ID1406970
'During cell division, sister chromosomes segregate from each other on a microtubule-based structure called the mitotic spindle. Proteins bind to the centromere, a region of chromosomal DNA, to form the kinetochore, which mediates chromosome attachment to the mitotic spindle microtubules. In the budding yeast Saccharomyces cerevisiae, genetic analysis has shown ... More
External Na dependence of ouabain-sensitive ATP:ADP exchange initiated by photolysis of intracellular caged-ATP in human red cell ghosts.
AuthorsKaplan JH, Hollis RJ
JournalNature
PubMed ID6255339
'Coupled active transport of Na+ and K+ across cellular plasma membranes is mediated by (Na+ + K+)-stimulated Mg2+-dependent ATPase. Active cation transport by this Na pump involves a cyclic Na-dependent phosphorylation of the enzyme by intracellular ATP and hydrolytic dephosphorylation of the phosphoenzyme, stimulated by K+ (ref. 1). In human ... More
K+-dependence of electrogenic transport by the NaK-ATPase.
AuthorsGropp T, Cornelius F, Fendler K
JournalBiochim Biophys Acta
PubMed ID9459597
'Charge translocation by the NaK-ATPase from shark rectal gland was measured by adsorption of proteoliposomes to a planar lipid membrane. The proteoliposomes were prepared by reconstitution of purified NaK-ATPase into liposomes consisting of E. coli lipids. The protein was activated by applying an ATP concentration jump produced by photolysis of ... More
The effects of MgADP on cross-bridge kinetics: a laser flash photolysis study of guinea-pig smooth muscle.
AuthorsNishiye E, Somlyo AV, Török K, Somlyo AP
JournalJ Physiol
PubMed ID8487195
'1. The effects of MgADP on cross-bridge kinetics were investigated using laser flash photolysis of caged ATP (P3-1(2-nitrophenyl) ethyladenosine 5'-triphosphate), in guinea-pig portal vein smooth muscle permeabilized with Staphylococcus aureus alpha-toxin. Isometric tension and in-phase stiffness transitions from rigor state were monitored upon photolysis of caged ATP. The estimated concentration ... More
Charge translocation by the Na,K-pump: I. Kinetics of local field changes studied by time-resolved fluorescence measurements.
AuthorsBühler R, Stürmer W, Apell HJ, Läuger P
JournalJ Membr Biol
PubMed ID1652643
'Membrane fragments containing a high density of Na,K-ATPase can be noncovalently labeled with amphiphilic styryl dyes (e.g., RH 421). Phosphorylation of the Na,K-ATPase by ATP in the presence of Na+ and in the absence of K+ leads to a large increase of the fluorescence of RH 421 (up to 100%). ... More
The role of ATP in the regulation of intracellular Ca2+ release in single fibres of mouse skeletal muscle.
AuthorsAllen DG, Lännergren J, Westerblad H
JournalJ Physiol
PubMed ID9051572
'1. Single fibres were dissected from mouse flexor brevis muscle and injected with indo-1 and the P3-1 (2-nitrophenyl)ethyl ester of ATP (caged ATP). Myoplasmic calcium concentration ([Ca2+]i) and force were monitored during single tetani or tetani repeated until force was reduced to about 30% of control values. In vitro experiments ... More
Distance of myofilament sliding per ATP molecule in skeletal muscle fibers studied using laser flash photolysis of caged ATP.
AuthorsYamada T, Abe O, Kobayashi T, Sugi H
JournalAdv Exp Med Biol
PubMed ID8109363
'We studied the distance of myofilament sliding per hydrolysis of one ATP molecule by recording shortening of single glycerinated muscle fibers induced by laser flash photolysis of caged ATP, diffusion of photochemically released ATP out of the fiber being prevented by surrounding the fiber with silicone oil. With 75 microM ... More
Flash photolysis studies of excitation-contraction coupling, regulation, and contraction in smooth muscle.
AuthorsSomlyo AP, Somlyo AV
JournalAnnu Rev Physiol
PubMed ID2184779
'1. Flash photolysis of caged compounds of phenylephrine, inositol 1, 4, 5 trisphosphate (InsP3), GTP gamma S, ATP, and CTP has been successfully used to study excitation-contraction coupling, contractile regulation, and contraction in smooth muscle. Major processes explored with this method were (a) the delay between agonist-receptor interaction and contraction ... More
Properties and uses of photoreactive caged compounds.
AuthorsMcCray JA, Trentham DR
JournalAnnu Rev Biophys Biophys Chem
PubMed ID2660825
The efficiency of contraction in rabbit skeletal muscle fibres, determined from the rate of release of inorganic phosphate.
AuthorsHe ZH, Chillingworth RK, Brune M, Corrie JE, Webb MR, Ferenczi MA
JournalJ Physiol
PubMed ID10358123
'1. The relationship between mechanical power output and the rate of ATP hydrolysis was investigated in segments of permeabilized fibres isolated from rabbit psoas muscle. 2. Contractions were elicited at 12 degrees C by photolytic release of ATP from the P3 -1-(2-nitrophenyl) ester of ATP (NPE-caged ATP). Inorganic phosphate (Pi) ... More
Correlation between mechanical and enzymatic events in contracting skeletal muscle fiber.
AuthorsShepard A, Borejdo J
JournalBiochemistry
PubMed ID15005615
'The conventional hypothesis of muscle contraction postulates that the interaction between actin and myosin involves tight coupling between the power stroke and hydrolysis of ATP. However, some in vitro experiments suggested that hydrolysis of a single molecule of ATP caused multiple mechanical cycles. To test whether the tight coupling is ... More
Photoreaction of caged ATP studied by the time-resolved transient grating method.
AuthorsChoi J, Terazima M
JournalPhotochem Photobiol Sci
PubMed ID12911225
'The photoreaction of caged ATP, P3-[1-(2-nitrophenyl)ethyl]adenosine-5''-triphosphate, has been investigated using the time-resolved transient grating (TG) method. We found that a feature of the TG signal time profile depends sensitively on the grating wavenumber (q) after the photoexcitation of caged ATP. This q-dependent feature of the TG signal was interpreted based ... More
Tension relaxation induced by pulse photolysis of caged ATP in partially crosslinked fibers from rabbit psoas muscle.
AuthorsEmoto Y, Horiuti K, Tawada K, Yamada K
JournalProc Natl Acad Sci U S A
PubMed ID7878001
'Muscle contractile force is thought to be generated by ATP-induced conformational changes in myosin crossbridges. In the present study, we investigated the response to ATP binding of force-bearing, attached cross-bridges. For this investigation, skinned fibers, in which myosin heads were in part covalently crosslinked to thin filaments with a zero-length ... More
Kinetics of force generation by single kinesin molecules activated by laser photolysis of caged ATP.
AuthorsHiguchi H, Muto E, Inoue Y, Yanagida T
JournalProc Natl Acad Sci U S A
PubMed ID9114000
'To relate transients of force by single kinesin molecules with the elementary steps of the ATPase cycle, we measured the time to force generation by kinesin after photorelease of ATP from caged ATP. Kinesin-coated beads were trapped by an infrared laser and brought onto microtubules fixed to a coverslip. Tension ... More
Photochemical reaction mechanisms of 2-nitrobenzyl compounds: methyl ethers and caged ATP.
AuthorsIl'ichev YV, Schwörer MA, Wirz J
JournalJ Am Chem Soc
PubMed ID15070376
'The mechanism of methanol photorelease from 2-nitrobenzyl methyl ether (1) and 1-(2-nitrophenyl)ethyl methyl ether (2), and of ATP release from adenosine-5''-triphosphate-[P(3)-(1-(2-nitrophenyl)ethyl)] ester (''caged ATP'', 3) was studied in various solvents by laser flash photolysis with UV-vis and IR detection. In addition to the well-known primary aci-nitro transients (A, lambda(max) approximately ... More
Motile properties of the kinesin-related Cin8p spindle motor extracted from Saccharomyces cerevisiae cells.
AuthorsGheber L, Kuo SC, Hoyt MA
JournalJ Biol Chem
PubMed ID10092642
'We have developed microtubule binding and motility assays for Cin8p, a kinesin-related mitotic spindle motor protein from Saccharomyces cerevisiae. The methods examine Cin8p rapidly purified from crude yeast cell extracts. We created a recombinant form of CIN8 that fused the biotin carrying polypeptide from yeast pyruvate carboxylase to the carboxyl ... More
Phosphate release and force generation in skeletal muscle fibers.
AuthorsHibberd MG, Dantzig JA, Trentham DR, Goldman YE
JournalScience
PubMed ID3159090
'Rapid laser pulse-induced photolysis of an adenosine triphosphate precursor in muscle fibers abruptly initiated cycling of the cross-bridges. The accompanying changes in tension and stiffness were related to elementary mechanochemical events of the energy-transducing mechanism. When inorganic phosphate was present at millimolar concentrations during liberation of adenosine triphosphate in the ... More
Transient contraction of muscle fibers on photorelease of ATP at intermediate concentrations of Ca2+.
AuthorsHoriuti K, Kagawa K, Yamada K
JournalBiophys J
PubMed ID7858129
'We isometrically activated skinned fibers in rigor by flash photolysis of caged ATP at various [Ca2+] at 8 degrees C. On release of ATP, tension initially decreased with the same time course at all [Ca2+]. At high [Ca2+] (pCa < or = 5.8), tension rose to the steady-state plateau after ... More
Laser-induced photogeneration of ATP: a new approach to the study of chemical kinetics of muscle contraction.
AuthorsHibberd MG, Goldman YE, Trentham DR
JournalCurr Top Cell Regul
PubMed ID6499523
Rapid photolytic release of adenosine 5'-triphosphate from a protected analogue: utilization by the Na:K pump of human red blood cell ghosts.
AuthorsKaplan JH, Forbush B, Hoffman JF
JournalBiochemistry
PubMed ID148906
Caged compounds and striated muscle contraction.
AuthorsHomsher E, Millar NC
JournalAnnu Rev Physiol
PubMed ID2139560
Temperature dependence of ATP release from "caged" ATP.
AuthorsBarabás K, Keszthelyi L
JournalActa Biochim Biophys Acad Sci Hung
PubMed ID6545637
The ATP release from "caged" ATP by laser flash was determined at different temperatures and pH values. We measured the time resolved absorbance changes of the coloured intermediate, which is involved in the photolytic process. The linearity of the Arrhenius curves suppose only one process and the calculated activation enthalpies ... More
Relaxation of muscle fibres by photolysis of caged ATP.
AuthorsGoldman YE, Hibberd MG, McCray JA, Trentham DR
JournalNature
PubMed ID7177194
A novel method has been developed for studying the reaction kinetics of the force-generating mechanism in muscle. Inert photolabile precursors of ATP or ADP are incorporated into muscle fibres having their surface membrane barrier removed. The nucleotide is then rapidly liberated by laser pulse photolysis. This circumvents the limitation in ... More
Kinetics of pump currents generated by the Na+,K+-ATPase.
AuthorsFendler K, Grell E, Bamberg E
JournalFEBS Lett
PubMed ID2824243
Purified Na+,K+-ATPase from pig kidney was attached to black lipid membranes. Pump currents of the enzyme could be measured with a time resolution of approx. 1 ms by releasing ATP from caged ATP with a UV laser flash. Analysis of the transient currents shows that a slow non-electrogenic step is ... More
Substrate binding and enzyme function investigated by infrared spectroscopy.
AuthorsBarth A, Zscherp C
JournalFEBS Lett
PubMed ID10908712
Protein conformational changes triggered by molecule binding are increasingly investigated by infrared spectroscopy often using caged compounds. Several examples of molecule-protein recognition studies are given, which focus on nucleotide binding to proteins. The investigation of enzyme mechanisms is illustrated in detail using the Ca(2+)-ATPase of the sarcoplasmic reticulum membrane as ... More
A low cost high intensity flash device for photolysis experiments.
AuthorsRapp G, Güth K
JournalPflugers Arch
PubMed ID3357758
Novel techniques of flash photolysis experiments require high intensity light sources in the near UV. We describe here a simple and inexpensive flash device which may complete with bulky and expensive laser systems if the experiments do not necessitate very short light pulses. Using a particular optical arrangement and stored ... More
Caged ATP - an internal calibration method for ATP bioluminescence assays.
ATP bioluminescence, based on the firefly luciferase system, is used for the rapid determination of hygienic practices in the food industry. This study has demonstrated the use of caged ATP as an internal ATP standard and quantified the effects of industrial cleansing solutions, alcoholic beverages and pH on firefly luciferase ... More
Comparison of nitrophenylethyl and hydroxyphenacyl caging groups.
AuthorsDu X, Frei H, Kim SH
JournalBiopolymers
PubMed ID11343283
Nitrophenylethyl (NPE)- and hydroxyphenacyl (HPA)-caged nucleotides were employed in a time-resolved Fourier transform IR spectroscopy study on Ras-catalyzed guanosine triphosphate (GTP) hydrolysis. A fast kinetic component was observed following the photolysis of NPE-caged nucleotides in the NPE-GTP-Ras complex. However, this kinetic component was not observed in the HPA-GTP-Ras experiment. This ... More
Caged compounds: photorelease technology for control of cellular chemistry and physiology.
AuthorsEllis-Davies GC
JournalNat Methods
PubMed ID17664946
Caged compounds are light-sensitive probes that functionally encapsulate biomolecules in an inactive form. Irradiation liberates the trapped molecule, permitting targeted perturbation of a biological process. Uncaging technology and fluorescence microscopy are 'optically orthogonal': the former allows control, and the latter, observation of cellular function. Used in conjunction with other technologies ... More
Reaction-induced infrared difference spectroscopy for the study of protein reaction mechanisms.
AuthorsZscherp C, Barth A
JournalBiochemistry
PubMed ID11329252
This paper reviews state-of-the-art reaction-induced infrared difference spectroscopy of proteins. This technique enables detailed characterization of enzyme function on the level of single bonds of proteins, cofactors, or substrates. The following methods to initiate a reaction in the infrared sample are discussed: (i) light-induced difference spectroscopy, (ii) attenuated total reflection ... More
Na+,K(+)-ATPase pump currents activated by an ATP concentration jump. Comparison of studies with purified membrane fragments and giant excised patches.
AuthorsNagel G, Friedrich T, Fendler K, Bamberg E
JournalAnn N Y Acad Sci
PubMed ID9405814
The lipid bilayer and the giant patch technique were used to study the Na+,K(+)-ATPase. In excised patches from ventricular myocytes, Na+,K(+)-pump currents show a saturable ATP dependence with a Km of approximately 150 microM at 24 degrees C. Partial reactions in the transport cycle were investigated by generating ATP concentration ... More
Inositol trisphosphate releases stored calcium to block voltage-dependent calcium channels in single smooth muscle cells.
AuthorsKomori S, Bolton TB
JournalPflugers Arch
PubMed ID1653941
In single cells obtained by enzymic treatment of rabbit small-intestinal smooth muscle, and held under voltage clamp by patch pipette in the whole-cell recording mode, release of inositol trisphosphate (InsP3) from its caged precursor by flash photolysis caused complete inhibition of the voltage-dependent calcium current. No inhibition was seen in ... More
The cross-bridge mechanism studied by flash photolysis of caged ATP in skeletal muscle fibers.
AuthorsHoriuti K
JournalJpn J Physiol
PubMed ID9504128
Flash photolysis of caged ATP was first employed in muscle cell physiology about 15 years ago. This technique enables examination of the millisecond dynamics of synchronized cross-bridge ATPases in muscle fibers, and consequently, many photolysis studies have been performed in attempt to elucidate the cross-bridge mechanism. The results of early ... More
Static and time-resolved structural studies of the Ca2+-ATPase of isolated sarcoplasmic reticulum.
AuthorsBlasie JK, Herbette L, Pierce D, Pascolini D, Scarpa A, Fleischer S
JournalAnn N Y Acad Sci
PubMed ID6220651
X-ray and neutron diffraction studies of oriented multilayers of isolated light sarcoplasmic reticulum (SR) have provided the separate profile structures of the lipid bilayer and the Ca2+-ATPase molecule within the membrane profile to approximately 10 A resolution. These studies utilized biosynthetically deuterated SR phospholipids incorporated isomorphously into the isolated SR ... More
Kinetics and mechanism of DNA repair. Evaluation of caged compounds for use in studies of u.v.-induced DNA repair.
AuthorsMeldrum RA, Shall S, Wharton CW
JournalBiochem J
PubMed ID2327973
Experiments are described in which the feasibility of using caged dideoxy and other nucleoside triphosphate analogues for trapping breaks induced by u.v. radiation damage to mammalian cell DNA is evaluated. These nucleotide analogues that have a photolabile 1-(2-nitrophenyl)ethyl-protecting group attached to the gamma-phosphate are placed in situ by permeabilizing cells ... More
Inhibition of unloaded shortening velocity in permeabilized muscle fibres by caged ATP compounds.
AuthorsThirlwell H, Sleep JA, Ferenczi MA
JournalJ Muscle Res Cell Motil
PubMed ID7622628
The effects of both the P3-1-(2-nitrophenyl)ethyl ester of adenosine 5'-triphosphate (NPE-caged ATP) and its separate diastereoisomers, and the P3-3',5'-dimethoxybenzoin ester of ATP (DMB-caged ATP) were studied on the unloaded shortening velocity of glycerinated rabbit psoas muscle fibres. The unloaded shortening velocities of the active fibres were measured as a function ... More
Effects of ionic strength on force transients induced by flash photolysis of caged ATP in covalently crosslinked rabbit psoas muscle fibers.
AuthorsYamada K, Emoto Y, Horiuti K, Tawada K
JournalAdv Exp Med Biol
PubMed ID8109361
Single fibers from glycerinated rabbit psoas muscle were treated with 1-ethyl-3[3-(dimethylamino) propyl] carbodiimide (EDC), after rigor was induced, to crosslink myosin heads to actin. The optimally pre-stretched (approximately 1.8%), partially crosslinked fibers produce a large force when MgATP is depleted, and this force is abolished when MgATP is reintroduced, even ... More
Inhibition of ATP binding to myofibrils and acto-myosin subfragment 1 by caged ATP.
AuthorsSleep J, Herrmann C, Barman T, Travers F
JournalBiochemistry
PubMed ID8193117
The inhibitory effect of P3-[1-(2-nitrophenyl)ethyl]adenosine 5'-triphosphate (caged ATP) on the binding of Mg2+-ATP to myofibrils was investigated. The most sensitive method was found to be the monitoring of single turnovers of [gamma-32P] ATP hydrolysis using the quench flow technique. The method was tested using ADP, which was found to have ... More
Modulation of ATP-sensitive potassium channel activity by flash-photolysis of 'caged-ATP' in rat heart cells.
AuthorsNichols CG, Niggli E, Lederer WJ
JournalPflugers Arch
PubMed ID2315010
We have used 'caged-ATP' to investigate the kinetic behavior of KATP channels in ventricular cells from rat heart. In whole cells, loaded with 'caged-ATP', an increase of intracellular [ATP] following a UV light flash produced a decrease of KATP channel current that was too slow (tau approximately 300 ms) to ... More
The effect of dicyclohexylcarbodiimide and cyclopiazonic acid on the difference FTIR spectra of sarcoplasmic reticulum induced by photolysis of caged-ATP and caged-Ca2+.
AuthorsBuchet R, Jona I, Martonosi A
JournalBiochim Biophys Acta
PubMed ID1532328
The photochemical release of Ca2+ from caged-Ca2+ in the absence of ATP, and the release of ATP from caged-ATP in the presence of Ca2+ induce characteristic difference FTIR spectra on rabbit sarcoplasmic reticulum that are related to the formation of Ca2-E1 and E approximately P intermediates of the Ca(2+)-ATPase, respectively. ... More
BDM compared with P(i) and low Ca2+ in the cross-bridge reaction initiated by flash photolysis of caged ATP.
AuthorsKagawa K, Horiuti K, Yamada K
JournalBiophys J
PubMed ID8599666
Using flash photolysis of caged ATP in skinned muscle fibers from rat psoas, we examined the inhibitory effects of 2,3-butanedione monoxime (BDM) on the contraction kinetics and the rate of ATP hydrolysis of the cross-bridges at approximately 10 degrees C. The hydrolysis rate was estimated from the stiffness records. The ... More
Ca2+ depletion and inositol 1,4,5-trisphosphate-evoked activation of Ca2+ entry in single guinea pig hepatocytes.
AuthorsGuihard G, Noel J, Capiod T
JournalJ Biol Chem
PubMed ID10788451
Store-operated Ca(2+) entry was investigated by monitoring the Ca(2+)-dependent K(+) permeability in voltage-clamped guinea pig hepatocytes. In physiological conditions, intracellular Ca(2+) stores are discharged following agonist stimulation, but depletion of this stores can be achieved using Ca(2+)-Mg(2+)-ATPase inhibitors such as 2,5-di(tert-butyl)-1,4-benzohydroquinone and thapsigargin. The effect of internal Ca(2+) store depletion ... More
Fluorescence polarization from isomers of tetramethylrhodamine at SH-1 in rabbit psoas muscle fibers.
AuthorsBerger CL, Craik JS, Trentham DR, Corrie JE, Goldman YE
JournalBiophys J
PubMed ID7787111
We have used fluorescence polarization to examine orientational changes of the 5- and 6-isomers of acetamidotetramethylrhodamine (ATR) covalently bound to SH-1 (Cys-707 of the myosin heavy chain) in single, skinned fibers from rabbit psoas muscle after rapid length steps or photolysis of caged nucleotides. Similar results were obtained with both ... More
Mitochondrial respiration and Ca2+ waves are linked during fertilization and meiosis completion.
AuthorsDumollard R, Hammar K, Porterfield M, Smith PJ, Cibert C, Rouvière C, Sardet C
JournalDevelopment
PubMed ID12505999
Fertilization increases both cytosolic Ca(2+) concentration and oxygen consumption in the egg but the relationship between these two phenomena remains largely obscure. We have measured mitochondrial oxygen consumption and the mitochondrial NADH concentration on single ascidian eggs and found that they increase in phase with each series of meiotic Ca(2+) ... More
A new approach to time-resolved studies of ATP-requiring biological systems; laser flash photolysis of caged ATP.
AuthorsMcCray JA, Herbette L, Kihara T, Trentham DR
JournalProc Natl Acad Sci U S A
PubMed ID6938971
2-Nitrobenzyl derivatives have been used for several years as photolabile protecting groups in synthetic organic chemistry. Recently, P3-1-(2-nitro phenylethyladenosine 5'-triphosphate "caged ATP" was synthesized and its photolysis was shown to generate ATP in situ. This and related reactions have great potential for structural and kinetic studies of both intact and ... More
Calcium pump kinetics determined in single erythrocyte ghosts by microphotolysis and confocal imaging.
AuthorsKubitscheck U, Pratsch L, Passow H, Peters R
JournalBiophys J
PubMed ID7669907
The activity of the plasma membrane calcium pump was measured in single cells. Human red blood cell ghosts were loaded with a fluorescent calcium indicator and either caged calcium and ATP (protocol A) or caged ATP and calcium (protocol B). In a suitably modified laser scanning microscope either calcium or ... More
Rate of phosphate release after photoliberation of adenosine 5'-triphosphate in slow and fast skeletal muscle fibers.
AuthorsHe Z, Stienen GJ, Barends JP, Ferenczi MA
JournalBiophys J
PubMed ID9788934
Inorganic phosphate (Pi) release was determined by means of a fluorescent Pi-probe in single permeabilized rabbit soleus and psoas muscle fibers. Measurements of Pi release followed photoliberation of approximately 1.5 mM ATP by flash photolysis of NPE-caged ATP in the absence and presence of Ca2+ at 15 degrees C. In ... More
Structural features of nonpeptide prenyl pyrophosphates that determine their antigenicity for human gamma delta T cells.
AuthorsMorita CT, Lee HK, Wang H, Li H, Mariuzza RA, Tanaka Y
JournalJ Immunol
PubMed ID11418629
Human Vgamma2Vdelta2(+) T cells proliferate in vivo during many microbial infections. We have found that Vgamma2Vdelta2(+) T cells recognize nonpeptide prenyl pyrophosphates and alkylamines. We now have defined structural features that determine the antigenicity of prenyl pyrophosphates by testing synthetic analogs for bioactivity. We find that the carbon chain closest ... More
Dynein-ADP as a force-generating intermediate revealed by a rapid reactivation of flagellar axoneme.
AuthorsTani T, Kamimura S
JournalBiophys J
PubMed ID10465762
Fragmented flagellar axonemes of sand dollar spermatozoa were reactivated by rapid photolysis of caged ATP. After a time lag of 10 ms, axonemes treated with protease started sliding disintegration. Axonemes without protease digestion started nanometer-scale high-frequency oscillation after a similar time lag. Force development in the sliding disintegration was measured ... More
Electroporation loading and photoactivation of caged InsP3: tools to investigate the relation between cellular ATP release in response to intracellular InsP3 elevation.
AuthorsBraet K, Mabilde C, Cabooter L, Rapp G, Leybaert L
JournalJ Neurosci Methods
PubMed ID14687677
Photolytic liberation of InsP(3) in single cells triggers cell-to-cell propagating calcium changes that are communicated by a gap junctional and a paracrine purinergic pathway involving InsP(3)-triggered ATP release. We investigated the relation between the InsP(3) stimulus and the resulting ATP release in ECV304 cells using UV photolysis of caged compounds ... More
Sub-second kinetics of the nitric oxide receptor, soluble guanylyl cyclase, in intact cerebellar cells.
AuthorsBellamy TC, Garthwaite J
JournalJ Biol Chem
PubMed ID11073946
Soluble guanylyl cyclase (sGC) catalyzes cGMP synthesis and serves as a physiological receptor for nitric oxide (NO). Recent evidence indicates that key properties of sGC within cells differ from those of purified sGC. We have devised a technique for resolving NO-stimulated sGC activity in cells on a sub-second time scale, ... More
Effect of cross-linking on the contractile behavior of myofibrils.
AuthorsHarrington WF, Karr T, Busa WB
JournalAdv Exp Med Biol
PubMed ID8109372
When rabbit psoas myofibrils in rigor are cross-linked with DMS (dimethyl suberimidate) for various periods of time, they contract on activation to a final sarcomere spacing of 1.3-1.5 microns. This behavior is observed out to 100 min cross-linking time (2 mg/ml DMS; 10 degrees C). Over the next 100 min ... More
Phosphoenzyme conversion of the sarcoplasmic reticulum Ca(2+)-ATPase. Molecular interpretation of infrared difference spectra.
AuthorsBarth A
JournalJ Biol Chem
PubMed ID10428781
Time-resolved Fourier transform infrared difference spectra of the phosphoenzyme conversion and Ca(2+) release reaction (Ca(2)E(1)-P --> E(2)-P) of the sarcoplasmic reticulum Ca(2+)-ATPase were recorded at pH 7 and 1 degrees C in H(2)O and (2)H(2)O. In the amide I spectral region, the spectra indicate backbone conformational changes preserving conformational changes ... More
Mechanical study of rat soleus muscle using caged ATP and X-ray diffraction: high ADP affinity of slow cross-bridges.
AuthorsHoriuti K, Yagi N, Takemori S
JournalJ Physiol
PubMed ID9263922
1. The cross-bridges in slow- and fast-twitch fibres (taken, respectively, from soleus and psoas muscles of rats) were examined in mechanical experiments using caged ATP and X-ray diffraction, to compare their binding of ATP and ADP. 2. Caged ATP was photolysed in rigor fibres. When ADP was removed from pre-photolysis ... More
Inositol 1,4,5-trisphosphate directs Ca(2+) flow between mitochondria and the Endoplasmic/Sarcoplasmic reticulum: a role in regulating cardiac autonomic Ca(2+) spiking.
AuthorsJaconi M, Bony C, Richards SM, Terzic A, Arnaudeau S, Vassort G, Pucéat M
JournalMol Biol Cell
PubMed ID10793156
The signaling role of the Ca(2+) releaser inositol 1,4, 5-trisphosphate (IP(3)) has been associated with diverse cell functions. Yet, the physiological significance of IP(3) in tissues that feature a ryanodine-sensitive sarcoplasmic reticulum has remained elusive. IP(3) generated by photolysis of caged IP(3) or by purinergic activation of phospholipase Cgamma slowed ... More
Interaction of a Dictyostelium member of the plastin/fimbrin family with actin filaments and actin-myosin complexes.
AuthorsPrassler J, Stocker S, Marriott G, Heidecker M, Kellermann J, Gerisch G
JournalMol Biol Cell
PubMed ID9017597
A protein purified from cytoskeletal fractions of Dictyostelium discoideum proved to be a member of the fimbrin/plastin family of actin-bundling proteins. Like other family members, this Ca(2+)-inhibited 67-kDa protein contains two EF hands followed by two actin-binding sites of the alpha-actinin/beta-spectrin type. Dd plastin interacted selectively with actin isoforms: it ... More
Effect of inorganic phosphate and ADP on the myofilament sliding induced by laser flash photolysis of caged ATP.
AuthorsWada H, Yamada T, Sugi H
JournalBiochim Biophys Acta
PubMed ID8664308
Using the technique of laser flash photolysis of caged ATP, we have suggested that, under nearly isometric conditions, the unitary distance of myofilament sliding per ATP molecule (myosin head powerstroke) is about 10 nm. To give further information about the mechanism of myofilament sliding, we studied the effect of inorganic ... More
Fast transient currents in Na,K-ATPase induced by ATP concentration jumps from the P3-[1-(3',5'-dimethoxyphenyl)-2-phenyl-2-oxo]ethyl ester of ATP.
AuthorsSokolov VS, Apell HJ, Corrie JE, Trentham DR
JournalBiophys J
PubMed ID9591656
Electrogenic ion transport by Na,K-ATPase was investigated by analysis of transient currents in a model system of protein-containing membrane fragments adsorbed to planar lipid bilayers. Sodium transport was triggered by ATP concentration jumps in which ATP was released from an inactive precursor by an intense near-UV light flash. The method ... More
Kinetics of relaxation from rigor of permeabilized fast-twitch skeletal fibers from the rabbit using a novel caged ATP and apyrase.
AuthorsThirlwell H, Corrie JE, Reid GP, Trentham DR, Ferenczi MA
JournalBiophys J
PubMed ID7696482
The complex time course of tension decay was investigated in fast-twitch permeabilized rabbit muscle fibers when they were relaxed from the rigor state using photochemical generation of ATP. A novel caged ATP compound, the P3-3',5'-dimethoxybenzoin ester of ATP (DMB-caged ATP), as well as the P3-1-(2-nitrophenyl)ethyl ester of ATP (NPE-caged ATP), ... More
Photoliberating inositol-1,4,5-trisphosphate triggers ATP release that is blocked by the connexin mimetic peptide gap 26.
AuthorsBraet K, Vandamme W, Martin PE, Evans WH, Leybaert L
JournalCell Calcium
PubMed ID12526886
Calcium signals can be communicated between cells by the diffusion of a second messenger through gap junction channels or by the release of an extracellular purinergic messenger. We investigated the contribution of these two pathways in endothelial cell lines by photoliberating InsP(3) or calcium from intracellular caged precursors, and recording ... More
Na+,K(+)-ATPase pump currents in giant excised patches activated by an ATP concentration jump.
AuthorsFriedrich T, Bamberg E, Nagel G
JournalBiophys J
PubMed ID8913588
The giant-patch technique was used to study the Na+,K(+)-ATPase in excised patches from rat or guinea pig ventricular myocytes. Na+,K(+)-pump currents showed a saturable ATP dependence with aK(m) of approximately 150 microM at 24 degrees C. The pump current can be completely abolished by ortho-vanadate. Dissociation of vanadate from the ... More
Effects of ethylene glycol on the kinetics of contraction on flash photolysis of caged ATP in rat psoas muscle fibres.
AuthorsHoriuti K, Sakoda T, Takei M, Yamada K
JournalJ Muscle Res Cell Motil
PubMed ID1597514
ATP (1-1.2 mM) was photoreleased from caged ATP (5 mM) in skinned fibres from rat psoas muscle at 15-17 degrees C, to examine the effects of ethylene glycol (EG; 20% in solvent) on the kinetics of isometric contraction. Muscle fibres were stretched by 0.5-2% before photolysis, so that force just ... More
Myosin essential light chain isoforms modulate the velocity of shortening propelled by nonphosphorylated cross-bridges.
AuthorsMatthew JD, Khromov AS, Trybus KM, Somlyo AP, Somlyo AV
JournalJ Biol Chem
PubMed ID9813037
The differential effects of essential light chain isoforms (LC17a and LC17b) on the mechanical properties of smooth muscle were determined by exchanging recombinant for endogenous LC17 in permeabilized smooth muscle treated with trifluoperazine (TFP). Co-precipitation with endogenous myosin heavy chain verified that 40-60% of endogenous LC17a could be exchanged for ... More
Infrared spectroscopic signals arising from ligand binding and conformational changes in the catalytic cycle of sarcoplasmic reticulum calcium ATPase.
AuthorsBarth A, Mäntele W, Kreutz W
JournalBiochim Biophys Acta
PubMed ID1826215
Fourier transform infrared spectroscopy was used to investigate ligand binding and conformational changes in the Ca2(+)-ATPase of sarcoplasmic reticulum during the catalytic cycle. The ATPase reaction was started in the infrared sample by release of ATP from the inactive, photolabile ATP derivative P3-1-(2-nitro)phenylethyladenosine 5'-triphosphate (caged ATP). Absorption spectroscopy in the ... More
Time-resolved charge movements in the sarcoplasmatic reticulum Ca-ATPase.
AuthorsPeinelt C, Apell HJ
JournalBiophys J
PubMed ID14747317
The time-resolved kinetics of the Ca(2+)-translocating partial reaction of the sarcoplasmatic reticulum Ca-ATPase was investigated by ATP-concentration jump experiments. ATP was released by an ultraviolet light flash from its inactive precursor and charge movements in the membrane domain of the ion pumps were detected by the fluorescent styryl dye 2BITC. ... More
Changes in orientation of actin during contraction of muscle.
AuthorsBorejdo J, Shepard A, Dumka D, Akopova I, Talent J, Malka A, Burghardt TP
JournalBiophys J
PubMed ID15041669
It is well documented that muscle contraction results from cyclic rotations of actin-bound myosin cross-bridges. The role of actin is hypothesized to be limited to accelerating phosphate release from myosin and to serving as a rigid substrate for cross-bridge rotations. To test this hypothesis, we have measured actin rotations during ... More
Ca2+-dependent K+ currents induced by muscarinic receptor activation in guinea pig adrenal chromaffin cells.
AuthorsOhta T, Ito S, Nakazato Y
JournalJ Neurochem
PubMed ID9489751
The characteristics of the outward current (I(out)) induced by muscarine were examined by using the whole-cell patch-clamp technique with a K+-containing pipette solution in combination with fura-2 microfluorometry in guinea pig chromaffin cells. Muscarine caused a transient increase in the cytosolic Ca2+ concentration ([Ca2+]i) and activated an I(out) with a ... More
Flash and smash: rapid freezing of muscle fibers activated by photolysis of caged ATP.
AuthorsHirose K, Lenart TD, Murray JM, Franzini-Armstrong C, Goldman YE
JournalBiophys J
PubMed ID8369445
A new approach was used to study transient structural states of cross-bridges during activation of muscle fibers. Rabbit skinned muscle fibers were rapidly and synchronously activated from the rigor state by photolysis of caged ATP in the presence of Ca2+. At several different times during the switch from rigor to ... More
Comparison of Na+/K(+)-ATPase pump currents activated by ATP concentration or voltage jumps.
AuthorsFriedrich T, Nagel G
JournalBiophys J
PubMed ID9199783
Using the giant patch technique, we combined two fast relaxation methods on excised patches from guinea pig cardiomyocytes to compare the rate constants of the involved reaction steps. Experiments were done in the absence of intra- or extracellular K+. Fast ATP concentration jumps were generated by photolysis of caged ATP ... More
Microsecond rotational dynamics of spin-labeled Ca-ATPase during enzymatic cycling initiated by photolysis of caged ATP.
AuthorsLewis SM, Thomas DD
JournalBiochemistry
PubMed ID1653011
We have measured the microsecond rotational motions of the sarcoplasmic reticulum (SR) Ca-ATPase as a function of enzyme-specific ligands, including those that induce active calcium transport. We labeled the Ca-ATPase with a maleimide spin probe and detected rotational dynamics using saturation-transfer electron paramagnetic resonance (ST-EPR). This probe's ST-EPR spectra have ... More
Fluorescence polarization of skeletal muscle fibers labeled with rhodamine isomers on the myosin heavy chain.
AuthorsBerger CL, Craik JS, Trentham DR, Corrie JE, Goldman YE
JournalBiophys J
PubMed ID8968602
Fluorescence polarization was used to examine orientational changes of Rhodamine probes in single, skinned muscle fibers from rabbit psoas muscle following either photolysis of caged nucleotides or rapid length changes. Fibers were extensively and predominantly labeled at SH1 (Cys-707) of the myosin heavy chain with either the 5- or the ... More
P(3)-[2-(4-hydroxyphenyl)-2-oxo]ethyl ATP for the rapid activation of the Na(+),K(+)-ATPase.
AuthorsGeibel S, Barth A, Amslinger S, Jung AH, Burzik C, Clarke RJ, Givens RS, Fendler K
JournalBiophys J
PubMed ID10968997
P(3)-[2-(4-hydroxyphenyl)-2-oxo]ethyl ATP (pHP-caged ATP) has been investigated for its application as a phototrigger for the rapid activation of electrogenic ion pumps. The yield of ATP after irradiation with a XeCl excimer laser (lambda = 308 nm) was determined at pH 6.0-7.5. For comparison, the photolytic yields of P(3)-[1-(2-nitrophenyl)]ethyl ATP (NPE-caged ... More
Relaxation of chemically skinned guinea pig taenia coli smooth muscle from rigor by photolytic release of adenosine-5'-triphosphate.
AuthorsArner A, Goody RS, Rapp G, Rüegg JC
JournalJ Muscle Res Cell Motil
PubMed ID3429640
The mechanical events following release of ATP from P3-1-(2-nitro)phenylethyladenosine-5'-triphosphate (caged-ATP) in skinned guinea pig taenia coli smooth muscle in rigor were investigated. A rigor force of about 25-35% of the maximal active force was obtained by removing ATP at the plateau of a maximal active contraction. In the rigor solution ... More
Pump currents generated by the purified Na+K+-ATPase from kidney on black lipid membranes.
AuthorsFendler K, Grell E, Haubs M, Bamberg E
JournalEMBO J
PubMed ID3004932
The transport activity of purified Na+K+-ATPase was investigated by measuring the electrical pump current induced on black lipid membranes. Discs containing purified Na+K+-ATPase from pig kidney were attached to planar lipid bilayers in a sandwich-like structure. After the addition of only microM concentrations of an inactive photolabile ATP derivative [P3-1-(2-nitro)phenylethyladenosine ... More