Synthesis and properties of a new fluorescent analog of ATP: adenosine-5'-triphosphoro-gamma-1-(5-sulfonic acid) napthylamidate.
AuthorsYarbrough LR
JournalBiochem Biophys Res Commun
PubMed ID656102
Synthesis and properties of adenosine-5'-triphosphoro-gamma-1-(5-sulfonic acid)naphthyl ethylamidate: a fluorescent nucleotide substrate for DNA-dependent RNA polymerase from Escherichia coli.
AuthorsWu FY, Abdulwajid AW, Solaiman D
JournalArch Biochem Biophys
PubMed ID2423030
'A new fluorescent ATP analog, adenosine-5''-triphosphoro-gamma-1-(5-sulfonic acid)naphthyl ethylamidate (gamma-1,5-EDANS)ATP, containing the fluorophore N-(aminoethyl)-5-naphthylamine-1-sulfonic acid attached via a gamma-phosphoamidate bond was synthesized in good yield. It has absorption maxima at 255 and 344 nm and a fluorescence emission maximum at 490 nm. These spectral characteristics permit its uses as an energy ... More
Proximity relationship between the active site of Escherichia coli RNA polymerase and rifampicin binding domain: a resonance energy-transfer study.
AuthorsKumar KP, Reddy PS, Chatterji D
JournalBiochemistry
PubMed ID1510938
'Escherichia coli RNA polymerase has two subsites, i and i + 1, for the binding of the first two substrates, and the first phosphodiester bond is formed between them during the initiation of transcription. Various studies have shown earlier that the inhibitor rifampicin has little effect, if any, on the ... More
Synthesis and characterization of fluorescent dinucleotide substrate for the DNA-dependent RNA polymerase from Escherichia coli.
AuthorsTyagi SC, Wu FY
JournalJ Biol Chem
PubMed ID2440871
'New fluorescent derivatives of dinucleoside monophosphates, (5''-AmNS)UpA/ApU/GpU/CpA, with a fluorophore, 1-aminonaphthalene-5-sulfonic acid (AmNS), attached to the first nucleotide of the dinucleoside monophosphates via a 5''-secondary amine linkage were synthesized in good yield. The chemical structure of (5''-AmNS)ApU was proved by the phosphodiesterase digestion followed by Whatman No. 3MM paper chromatographic ... More
Proximity between nucleotide/dinucleotide and metal ion binding sites in DNA-dependent RNA polymerase from Escherichia coli.
AuthorsTyagi SC, Wu FY
JournalBiochemistry
PubMed ID1321660
'In order to understand translocation in transcription, it is important to develop a continuous functional assay for RNA polymerase (RNAP) activity in vitro. Fluorescent derivatives of ATP, UTP, UpA, and CpA with aminonaphthalene-5-sulfonic acid (AmNS) attached to the nucleotide triphosphates via a gamma-phosphoramidate bond or to the dinucleotide monophosphates via ... More
Fluorescent nucleotide triphosphate substrates for snake venom phosphodiesterase.
AuthorsPollack SE, Auld DS
JournalAnal Biochem
PubMed ID6299131
Fluorescent nucleotide analogs: synthesis and applications.
AuthorsJameson DM, Eccleston JF
JournalMethods Enzymol
PubMed ID9170323
Spectroscopic techniques for study of phosphodiester bond formation by Escherichia coli RNA polymerase.
AuthorsSchlageck JG, Baughman M, Yarbrough LR
JournalJ Biol Chem
PubMed ID227888
Nucleotides containing the fluorophore 1-aminonaphthalene-5-sulfonate attached to the gamma phosphoryl group via a phosphoamidate bond are excellent substrates for Escherichia coli DNA-dependent RNA polymerase. Cleavage of the alpha-beta-phosphoryl bond produces significant changes in both absorption and fluorescence spectra. These alterations provide a sensitive and precise means for continuous monitoring of ... More
Synthesis and properties of fluorescent nucleotide substrates for DNA-dependent RNA polymerases.
AuthorsYarbrough LR, Schlageck JG, Baughman M
JournalJ Biol Chem
PubMed ID387781
A new class of fluorescent nucleotide analogs which contain the fluorophore 1-aminonaphthalene-5-sulfonate attached via a gamma-phosphoamidate bond has been synthesized. Both the purine and pyrimidine analogs have fluorescence emission maxima at 460 nm. Cleavage of the alpha-beta-phosphoryl bond produces change in both the absorption and fluorescence emission spectra. The fluorescence ... More
Stacking interactions in fluorescent nucleotide analogs containing 1-aminonaphthalene-5-sulfonate at the phosphoryl terminus.
AuthorsYarbrough LR, Bock JL
JournalJ Biol Chem
PubMed ID6253457
The conformational properties of nucleotides containing the fluorophore 1-aminonaphthalene-5-sulfonate attached via a gamma-phosphoamidate bond have been examined. Measurements of fluorescence excitation spectra show that energy absorbed by adenine in the ATP analog is transferred to the naphthalene moiety with an efficiency of approximately 43%. In nonpolar solvents transfer is almost ... More
Fluorescent nucleotides: application in a fluorescence immune assay.
AuthorsKardost RR, Voss EW
JournalMol Immunol
PubMed ID7043243
A fluorescence immune assay designed to measure anti-nucleotide antibody activity is described based on the synthesis of a fluorescent nucleotide probe possessing a low fluorescence quantum yield when free in aqueous solution (neutral pH). The fluorophore, AmNS (1-naphthylamine-5-sulfonic acid), was covalently conjugated to various phosphate derivatives of nucleotides through carbodiimide ... More
Titration calorimetric studies to elucidate the specificity of the interactions of polymyxin B with lipopolysaccharides and lipid A.
AuthorsSrimal S, Surolia N, Balasubramanian S, Surolia A
JournalBiochem J
PubMed ID8615847
Lipopolysaccharide (LPS), the major cell wall constituent of Gram-negative bacteria, evokes a multitude of biological effects in mammals including pyrogenicity and toxic shock syndrome. Polymyxin B (PmB), a polycationic cyclic peptide, is known to neutralize most of its activities. The nature of the interaction of PmB with LPS and lipid ... More
Fluorescence of intramolecular and intermolecular interactions of aminonaphthyl-sulfonate with nucleotides.
AuthorsFick J, Lawaczeck R, Schneider FW
JournalEur J Biochem
PubMed ID7128595
Fluorescence studies of the intramolecular and intermolecular interactions between aminonaphthylsulfonate and nucleotides of uracil or adenine are described. The fluorescence originates solely from the naphthyl moiety and is intramolecularly quenched by the base, uracil being more effective than adenine. The enzymatic splitting of the molecule into a nucleoside monophosphate and ... More
Mechanistic studies of reaction coupling in Glu-tRNAGln amidotransferase.
AuthorsHoriuchi KY, Harpel MR, Shen L, Luo Y, Rogers KC, Copeland RA
JournalBiochemistry
PubMed ID11371208
Organisms lacking Gln-tRNA synthetase produce Gln-tRNA(Gln) from misacylated Glu-tRNA(Gln) through the transamidation activity of Glu-tRNA(Gln) amidotransferase (Glu-AdT). Glu-AdT hydrolyzes Gln to Glu and NH(3), using the latter product to transamidate Glu-tRNA(Gln) in concert with ATP hydrolysis. In the absence of the amido acceptor, Glu-tRNA(Gln), the enzyme has basal glutaminase activity ... More
Fluorescence resonance energy transfer studies on the proximity relationship between the intrinsic metal ion and substrate binding sites of Escherichia coli RNA polymerase.
AuthorsWu FY, Tyagi SC
JournalJ Biol Chem
PubMed ID3308870
DNA-dependent RNA polymerase from Escherichia coli contains 2 mol of zinc/mol of holoenzyme (alpha 2 beta beta' sigma) with one zinc each in the beta and beta' subunits. A new method to substitute selectively the zinc in the beta subunit was developed by the inactivation of RNA polymerase with 0.25 ... More
Substrate-site interactions in the membrane-bound adenine-nucleotide carrier as disclosed by ADP and ATP analogs.
AuthorsBlock MR, Vignais PV
JournalBiochim Biophys Acta
PubMed ID6093873
The binding parameters of a number of ADP or ATP analogs to the adenine nucleotide carrier in mitochondria and inside-out submitochondrial particles have been explored by means of two specific inhibitors, carboxyatractyloside and bongkrekic acid. The nucleotides tested fell into two classes depending on the shape of the binding curve. ... More
Amino acid substitutions in the two largest subunits of Escherichia coli RNA polymerase that suppress a defective Rho termination factor affect different parts of the transcription complex.
AuthorsHeisler LM, Feng G, Jin DJ, Gross CA, Landick R
JournalJ Biol Chem
PubMed ID8662850
Among the earliest rpoBC mutations identified are three suppressors of the conditional lethal rho allele, rho201. These three mutations are of particular interest because, unlike rpoB8, they do not increase termination at all rho-dependent and rho-independent terminators. rpoB211 and rpoB212 both change Asn-1072 to His in conserved region H of ... More