Structural characteristics of the nucleotide-binding site of Escherichia coli primary replicative helicase DnaB protein. Studies with ribose and base-modified fluorescent nucleotide analogs.
AuthorsBujalowski W,Klonowska MM
JournalBiochemistry
PubMed ID8161526
Structural characteristics of the base- and ribose-binding regions of the high-affinity noninteracting nucleotide-binding site of Escherichia coli primary replicative helicase DnaB protein have been studied, using the base-modified fluorescent nucleotide analog 1, N6-ethenoadenosine diphosphate (epsilon ADP) and the ribose-modified fluorescent analogs 3'(2')-O-(N-methylantraniloyl)adenosine 5'-diphosphate (MANT-ADP), 3'-O-(N-methylantraniloyl)deoxyadenosine 5'-diphosphate (MANT-dADP), 3'-O-(N-methylantraniloyl)-deoxyadenosine 5'-triphosphate (MANT-dATP), ... More
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
Active site mutations in CheA, the signal-transducing protein kinase of the chemotaxis system in Escherichia coli.
AuthorsHirschman A, Boukhvalova M, VanBruggen R, Wolfe AJ, Stewart RC
JournalBiochemistry
PubMed ID11705377
'We investigated the functional roles of putative active site residues in Escherichia coli CheA by generating nine site-directed mutants, purifying the mutant proteins, and quantifying the effects of those mutations on autokinase activity and binding affinity for ATP. We designed these mutations to alter key positions in sequence motifs conserved ... 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
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
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
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
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
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
A subunit interaction in chloroplast ATP synthase determined by genetic complementation between chloroplast and bacterial ATP synthase genes.
AuthorsChen Z, Spies A, Hein R, Zhou X, Thomas BC, Richter ML, Gegenheimer P
JournalJ Biol Chem
PubMed ID7615507
'F1F0-ATP synthases utilize protein conformational changes induced by a transmembrane proton gradient to synthesize ATP. The allosteric cooperativity of these multisubunit enzymes presumably requires numerous protein-protein interactions within the enzyme complex. To correlate known in vitro changes in subunit structure with in vivo allosteric interactions, we introduced the beta subunit ... 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
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
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
Influence of divalent cations on nucleotide exchange and ATPase activity of chloroplast coupling factor 1.
AuthorsDigel JG, Moore ND, McCarty RE
JournalBiochemistry
PubMed ID9860834
'The ATPase activity of the catalytic part of ATP synthases is inhibited by free Mg2+, even though MgATP is the substrate. Here we show that the inhibition of the MgATPase activity of chloroplast coupling factor 1 deficient in its epsilon subunit (CF1-epsilon) by Mg2+ is complex. The hydrolysis of MgATP ... 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
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
Effect of covalent binding of a derivative of 2',3'-O-(2,4,6-trinitrophenyl)-ADP to the tight binding site of CF1 on the enzyme activity.
AuthorsHisabori T, Kothen G, Strotmann H
JournalJ Biochem (Tokyo)
PubMed ID8282720
'Irradiation of isolated chloroplast thylakoids with TNP-ADP results in non-covalent binding and covalent incorporation of a reaction product of TNP-ADP formed by photosynthetic reduction into the so-called "tight" nucleotide binding site of CF1 [Ponse et al. (1992) Z. Naturforsch. 47c, 264-274]. CF1 extracted from thus-loaded thylakoid membranes yielded maximal incorporation ... 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
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
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
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
The mechanism of inhibition of the Ca2+-ATPase by mastoparan. Mastoparan abolishes cooperative ca2+ binding.
AuthorsLongland CL, Mezna M, Michelangeli F
JournalJ Biol Chem
PubMed ID10329678
'The amphiphilic peptide mastoparan, isolated from wasp venom, is a potent inhibitor of the sarcoplasmic reticulum Ca2+-ATPase. At pH 7. 2, ATPase activity is inhibited with an inhibitory constant (Ki) of 1 +/- 0.13 microM. Mastoparan shifts the E2-E1 equilibrium toward E1 and may affect the regulatory ATP binding site. ... 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
Nucleotide binding of an ADP analog to cooperating sites of chloroplast F1-ATPase (CF1).
AuthorsGünther S, Huchzermeyer B
JournalEur J Biochem
PubMed ID10632724
'Pre-steady state nucleotide binding to the chloroplast F1-ATPase (CF1) was measured in a stopped-flow apparatus by monitoring the change of fluorescence intensity of TNP-ADP upon binding. The analysis of the time courses led to the proposal of a mechanism of nucleotide binding with the following characteristics. (a) It involves three ... 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
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
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
Nucleotide binding to IAF-labelled Na+/K(+)-ATPase measured by steady state fluorescence quenching by TNP-ADP.
AuthorsHellen EH, Pratap PR
JournalBiophys Chem
PubMed ID9474751
'Nucleotide binding to 5-iodoacetamidofluorescein (IAF) labelled Na+/K(+)-ATPase was measured by steady state fluorescence quenching of the fluorescein label via energy transfer to trinitrophenyl (TNP) labelled nucleotide. TNP-nucleotides are valuable probes of nucleotide binding to ATPases. Interpretation of these and other experiments in our laboratory using TNP-nucleotides with the Na+/K(+)-ATPase rely ... More
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
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
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
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
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
Using 2'(3')-O-trinitrophenyl derivatives of adenine nucleotides to study the structure and mechanism of functioning of soluble mitochondrial ATPase.
The 2'(3')-O-trinitrophenyl (N3ph) derivatives of the adenine nucleotides are strong competitive inhibitors of isolated mitochondrial ATPase (factor F1). Ki decreases in the order N3phAdo greater than N3phAdo greater than N3phAMP greater than N3phADP and is equal to 8 nM for N3phADP. Picric acid, which activates the ATPase reaction of factor ... More
A fluorescence investigation of the nucleotide binding sites of the Ca ATPase.
AuthorsWhite TE, Dewey TG
JournalMembr Biochem
PubMed ID2963204
Competitive binding and fluorescence energy transfer experiments were used to examine the binding of 2'[3']-O-(2,4,6-trinitrophenyl) adenosine-5'-diphosphate (TNP-ADP) to the catalytic site of Ca ATPase. Fluorescein isothiocyanate (FITC), which is known to covalently bind near the catalytic site (13), was shown to exclude all TNP-ADP binding. TNP-ADP, in turn, will protect ... More
Biological activities and spectroscopic properties of chromophoric and fluorescent analogs of adenine nucleoside and nucleotides, 2',3'-O-(2,4,6-trinitrocyclohexadienylidene) adenosine derivatives.
AuthorsHiratsuka T
JournalBiochim Biophys Acta
PubMed ID6295507
The ribose-modified chromophoric and fluorescent analog of ATP 2',3'-O-(2,4,6-trinitrocyclohexadienylidene) adenosine 5'-triphosphate (TNP-ATP) has been synthesized previously (Hiratsuka, T., and Uchida, K. (1973) Biochim. Biophys. Acta 320, 635-647 and Hiratsuka, T. (1976) Biochim. Biophys. Acta 453, 293-297). In the present study, four TNP-derivatives of ATP, ADP, AMP and adenosine were synthesized ... More
Structural mapping of nucleotide binding sites on chloroplast coupling factor.
AuthorsCerione RA, Hammes GG
JournalBiochemistry
PubMed ID6462173
Fluorescence resonance energy transfer was used to measure the distances between three nucleotide binding sites on solubilized chloroplast coupling factor from spinach and between each nucleotide site and two tyrosine residues which are important for catalytic activity. The nucleotide energy donor was 1,N6-ethenoadenosine di- or triphosphate, and the nucleotide energy ... More
Further investigations on the inorganic phosphate binding site of beef heart mitochondrial F1-ATPase.
AuthorsPougeois R, Lauquin GJ
JournalBiochemistry
PubMed ID2859884
The possibility that 4-azido-2-nitrophenyl phosphate (ANPP), a photoreactive derivative of inorganic phosphate (Pi) [Lauquin, G., Pougeois, R., & Vignais, P. V. (1980) Biochemistry 19, 4620-4626], could mimic ATP was investigated. ANPP was hydrolyzed in the dark by sarcoplasmic reticulum Ca2+-ATPase in the presence of Ca2+ but not in the presence ... More
Engineering ATPase activity in the isolated ABC cassette of human TAP1.
AuthorsErnst R, Koch J, Horn C, Tampé R, Schmitt L
JournalJ Biol Chem
PubMed ID16864587
The human transporter associated with antigen processing (TAP) translocates antigenic peptides from the cytosol into the endoplasmic reticulum lumen. The functional unit of TAP is a heterodimer composed of the TAP1 and TAP2 subunits, both of which are members of the ABC-transporter family. ABC-transporters are ATP-dependent pumps, channels, or receptors ... More
Distance measurement between the active site and cysteine-177 of the alkali one light chain of subfragment 1 from rabbit skeletal muscle.
AuthorsMoss DJ, Trentham DR
JournalBiochemistry
PubMed ID6140026
Förster energy-transfer techniques have been applied to labeled myosin subfragment 1 from rabbit skeletal muscle to determine an intramolecular distance and whether this distance changes during magnesium-dependent ATPase activity. The alkali one light chain was labeled at Cys-177 with N-(iodoacetyl)-N'-(5-sulfo-1-naphthyl)ethylenediamine (1,5-IAEDANS) and then exchanged into subfragment 1. High specificity of ... More
Expression and functional properties of the second predicted nucleotide binding fold of the cystic fibrosis transmembrane conductance regulator fused to glutathione-S-transferase.
AuthorsRandak C, Roscher AA, Hadorn HB, Assfalg-Machleidt I, Auerswald EA, Machleidt W
JournalFEBS Lett
PubMed ID7537226
CFTR-NBF-2 was expressed in Escherichia coli in fusion with glutathione-S-transferase, the soluble portion was purified and identified as a structured protein by its CD spectrum. Association reactions of the recombinant NBF-2 with adenine nucleotides were monitored qualitatively by demonstrating its ability to bind specifically to ATP-, ADP- and AMP-affinity agarose ... More
Two tight binding sites for ADP and their interactions during nucleotide exchange in chloroplast coupling factor 1.
AuthorsDigel JG, McCarty RE
JournalBiochemistry
PubMed ID7578053
Chloroplast coupling factor 1 (CF1) deficient in its epsilon subunit was loaded with 2'(3')-O-trinitrophenyl-ADP (TNP-ADP), and the release of tightly bound TNP-ADP was followed as a decrease in fluorescence. TNP-ADP could be exchanged for medium ADP, ATP, MgADP, and MgATP. The preferred substrate for exchange was MgADP, particularly in the ... More
Differences between two tight ADP binding sites of the chloroplast coupling factor 1 and their effects on ATPase activity.
AuthorsDigel JG, Kishinevsky A, Ong AM, McCarty RE
JournalJ Biol Chem
PubMed ID8702714
Purified chloroplast ATP synthase (CF1) contains 1.2-2 mol of tightly bound ADP/mol of enzyme that resists removal by gel filtration or dialysis. CF1 was depleted of its endogenous nucleotide by treatment with alkaline phosphatase. Tightly bound nucleotide was demonstrated not to have an essential structural role. CF1 depleted of endogenous ... More
Sequential hydrolysis of ATP molecules bound in interacting catalytic sites of Escherichia coli transcription termination protein Rho.
AuthorsStitt BL, Xu Y
JournalJ Biol Chem
PubMed ID9756883
Escherichia coli transcription termination protein Rho, an RNA-dependent ATPase, disrupts transcription complexes, releasing RNA and allowing RNA polymerase to recycle. Homohexameric Rho binds three molecules of MgATP in a single class of catalytically competent sites. In rapid mix chemical quench experiments, when Rho saturated with ATP was mixed with RNA ... More
Cystic fibrosis transmembrane conductance regulator: nucleotide binding to a synthetic peptide.
Multiple mutations in the gene responsible for cystic fibrosis are located within a region predicted to encode a nucleotide-binding fold in the amino terminal half of the cystic fibrosis transmembrane conductance regulator protein. A 67-amino acid peptide (P-67) that corresponds to the central region of this putative nucleotide binding site ... More
ATPase activity of a highly stable alpha(3)beta(3)gamma subcomplex of thermophilic F(1) can be regulated by the introduced regulatory region of gamma subunit of chloroplast F(1).
AuthorsBald D, Noji H, Stumpp MT, Yoshida M, Hisabori T
JournalJ Biol Chem
PubMed ID10777572
A mutant F(1)-ATPase alpha(3)beta(3)gamma subcomplex from the thermophilic Bacillus PS3 was constructed, in which 111 amino acid residues (Val(92) to Phe(202)) from the central region of the gamma subunit were replaced by the 148 amino acid residues of the homologous region from spinach chloroplast F(1)-ATPase gamma subunit, including the regulatory ... More
Catalytic activities of alpha3beta3gamma complexes of F1-ATPase with 1, 2, or 3 incompetent catalytic sites.
AuthorsAmano T, Hisabori T, Muneyuki E, Yoshida M
JournalJ Biol Chem
PubMed ID8663463
In order to know how many functional catalytic sites are necessary for ATPase activity of F1-ATPase from a thermophilic Bacillus PS3, a new method of isolating homogeneous preparations of the alpha3beta3gamma complex with 1, 2, or 3 incompetent catalytic sites was developed. Ten glutamic acids (Glu.Tag) were linked to the ... More
Large scale purification of detergent-soluble P-glycoprotein from Pichia pastoris cells and characterization of nucleotide binding properties of wild-type, Walker A, and Walker B mutant proteins.
AuthorsLerner-Marmarosh N, Gimi K, Urbatsch IL, Gros P, Senior AE
JournalJ Biol Chem
PubMed ID10574938
P-glycoprotein (Pgp; mouse MDR3) was expressed in Pichia pastoris, grown in fermentor culture, and purified. The final pure product is of high specific ATPase activity and is soluble at low detergent concentration. 120 g of cells yielded 6 mg of pure Pgp; >4 kg of cells were obtained from a ... More
Close proximity of tryptophan residues and ATP-binding site in Escherichia coli primary replicative helicase DnaB protein. Molecular topography of the enzyme.
AuthorsBujalowski W, Klonowska MM
JournalJ Biol Chem
PubMed ID7989300
The binding of fluorescent nucleotide analogs to the Escherichia coli primary replicative helicase DnaB protein causes strong quenching of protein tryptophan fluorescence. It results from the efficient fluorescence energy transfer (E) from tryptophans to analogs bound in the nucleotide-binding site, indicating that protein tryptophans are "clustered" in close proximity to ... More
Cross-linking of two beta subunits in the closed conformation in F1-ATPase.
AuthorsTsunoda SP, Muneyuki E, Amano T, Yoshida M, Noji H
JournalJ Biol Chem
PubMed ID10026189
In the crystal structure of mitochondrial F1-ATPase, two beta subunits with a bound Mg-nucleotide are in "closed" conformations, whereas the third beta subunit without bound nucleotide is in an "open" conformation. In this "CCO" (beta-closed beta-closed beta-open) conformational state, Ile-390s of the two closed beta subunits, even though they are ... More
Structural mapping of catalytic site with respect to alpha-subunit and noncatalytic site in yeast mitochondrial F1-ATPase using fluorescence resonance energy transfer.
AuthorsDivita G, Goody RS, Gautheron DC, Di Pietro A
JournalJ Biol Chem
PubMed ID8514756
The intrinsic tryptophan fluorescence of Schizosaccharomyces pombe mitochondrial F1 is a very sensitive probe to differentiate nucleotide binding to catalytic and noncatalytic sites (Divita, G., Di Pietro, A., Roux, B., and Gautheron, D. C. (1992) Biochemistry 31, 5791-5798), the catalytic site saturation producing quenching of Trp-257 fluorescence (Divita, G., Jault, ... More
Interaction of mitochondrial F1-ATPase with trinitrophenyl derivatives of ATP and ADP. Participation of third catalytic site and role of Mg2+ in enzyme inactivation.
AuthorsMurataliev MB, Boyer PD
JournalJ Biol Chem
PubMed ID8195184
Relatively high ATP concentrations show an unexpected lack of inhibition of the hydrolysis of low concentrations of trinitrophenyl ATP (TNP-ATP) by mitochondrial F1-ATPase. In striking contrast low TNP-ATP concentrations markedly inhibit the hydrolysis of much higher ATP concentrations. The three catalytic sites undergoing sequential conformational changes have different conformations at ... More
Overexpression and purification of the carboxyl-terminal nucleotide-binding domain from mouse P-glycoprotein. Strategic location of a tryptophan residue.
AuthorsBaubichon-Cortay H, Baggetto LG, Dayan G, Di Pietro A
JournalJ Biol Chem
PubMed ID7916013
The cDNA encoding the C-terminal nucleotide-binding domain (NBD2) from mouse P-glycoprotein involved in multidrug resistance was obtained from adrenal cell mRNA and amplified by reverse transcriptase polymerase chain reaction. NBD2 was highly overexpressed in Escherichia coli in fusion with glutathione S-transferase and could be purified after efficient thrombin cleavage. Both ... More
Glucose phosphorylation. Interaction of a 50-amino acid peptide of yeast hexokinase with trinitrophenyl ATP.
AuthorsArora KK, Shenbagamurthi P, Fanciulli M, Pedersen PL
JournalJ Biol Chem
PubMed ID2318895
A 50-amino acid peptide predicted by chemical modification studies of yeast hexokinase to contain an ATP-binding site has been synthesized and purified. The peptide, which includes residues from glutamate 78 at the NH2-terminal end to leucine 127 at the COOH-terminal, resides within the smaller of the two lobes found in ... More
The presence of two hydrolytic sites on beef heart mitochondrial adenosine triphosphatase.
AuthorsGrubmeyer C, Penefsky HS
JournalJ Biol Chem
PubMed ID6452454
The ribose-modified nucleotides 2',3'-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate (TNP-ATP) and TNP-ADP were used to probe the catalytic sites on soluble beef heart mitochondrial adenosine triphosphatase (F1). Both compounds were potent competitive inhibitors of ATP hydrolysis catalyzed by F1, Ki = 5.5 and 10 nM, respectively, and by submitochondrial particles, Ki (TNP-ATP) = ... More
Regulation of cytochrome c oxidase by interaction of ATP at two binding sites, one on subunit VIa.
AuthorsTaanman JW, Turina P, Capaldi RA
JournalBiochemistry
PubMed ID7918401
Cytochrome c oxidase isolated from a wild-type yeast strain and a mutant in which the gene for subunit VIa had been disrupted were used to study the interaction of adenine nucleotides with the enzyme complex. At low ionic strength (25 mM potassium phosphate), in the absence of nucleotides, the cytochrome ... More
Interaction of high-affinity nucleotide binding sites in mitochondrial ATP synthesis and hydrolysis.
AuthorsSchäfer G, Weber J
JournalJ Bioenerg Biomembr
PubMed ID6219105
The present study contributes to the problem of the dynamic structure of mitochondrial F1-ATPase and the functional interrelation of so-called tight nucleotide binding sites. Nucleotide analogs are used as a tool to differentiate two distinct functional states of the membrane-bound enzyme, proposed to reflect corresponding conformational states; they reveal F1-ATPase ... More
Ligands presumed to label high affinity and low affinity ATP binding sites do not interact in an (alpha beta)2 diprotomer in duck nasal gland Na+,K+-ATPase, nor Do the sites coexist in native enzyme.
AuthorsMartin DW, Sachs JR
JournalJ Biol Chem
PubMed ID10831595
The interaction of ligands deemed to be ATP analogues with renal Na(+),K(+)-ATPase suggests that two ATP binding sites coexist on each functional unit. Previous studies in which fluorescein 5-isothiocyanate (FITC) was used to label the high affinity ATP site and 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-diphosphate (TNP-ADP) was used to probe the low affinity ... More
Ca2+-induced distance change between points on actin and troponin in skeletal muscle thin filaments estimated by fluorescence energy transfer spectroscopy.
AuthorsMiki M, Kobayashi T, Kimura H, Hagiwara A, Hai H, Maéda Y
JournalJ Biochem (Tokyo)
PubMed ID9538210
Fluorescence resonance energy transfer spectroscopy has been used to study the spatial relationships between probes attached to actin and troponin in the reconstituted skeletal muscle thin filament in the presence and absence of Ca2+ ions. Gln-41 and the nucleotide-binding site of actin were selectively labeled with the acceptor probe: fluorescein ... More
Stoichiometry and affinity of nucleotide binding to P-glycoprotein during the catalytic cycle.
AuthorsQu Q, Russell PL, Sharom FJ
JournalBiochemistry
PubMed ID12549939
Drug transport mediated by P-glycoprotein (Pgp) is driven by hydrolysis of ATP at the two cytosolic nucleotide binding domains. However, little is currently known concerning the stoichiometry of nucleotide binding and how both stoichiometry and binding affinity change during the catalytic cycle of the transporter. To address this issue, we ... More
Two distinct classes of nucleotide binding sites in sarcoplasmic reticulum Ca-ATPase revealed by 2',3'-O-(2,4,6-trinitrocyclohexadienylidene)-ATP.
AuthorsDupont Y, Pougeois R, Ronjat M, Verjovsky-Almeida S
JournalJ Biol Chem
PubMed ID3158655
It was previously reported that 2',3'-O-(2,4,6-trinitrocyclohexadienylidene) (TNP)-nucleotides bind with high affinity to the sarcoplasmic reticulum Ca-ATPase (Dupont, Y., Chapron, Y., and Pougeois, R. (1982) Biochem. Biophys. Res. Commun. 106, 1272-1279 and Watanabe, T., and Inesi, G. (1982) J. Biol. Chem. 257, 11510-11516). Here we report a study of the Ca-ATPase ... More
Fluorescence energy transfer between probes on actin and probes on myosin.
AuthorsDos Remedios CG, Cooke R
JournalBiochim Biophys Acta
PubMed ID6743667
The structural relationship between F-actin filaments and the biologically active fragments of myosin (either as myosin subfragment-1 or heavy meromyosin) has been investigated using the technique of fluorescence energy transfer. Donor and acceptor probes were used to obtain the following inter- and intramolecular distances. Energy transfer was measured: (1) from ... More
Affinity labeling of two nucleotide sites on Na,K-ATPase using 2'(3')-O-(2,4,6-trinitrophenyl)8-azidoadenosine 5'-[alpha-32P]diphosphate (TNP-8N3-[alpha-32P]ADP) as a photoactivatable probe. Label incorporation before and after blocking the high affinity ATP site with fluorescein isothiocyanate.
AuthorsWard DG, Cavieres JD
JournalJ Biol Chem
PubMed ID9837964
ATP and its analogues act on the minimal functional unit of Na, K-ATPase, the alpha beta protomer, with high and low affinity effects. Fluorescein isothiocyanate (FITC) irreversibly blocks the high affinity, or catalytic, ATP site, and yet the surviving K+-phosphatase activity of soluble FITC-modified alphabeta protomers can be photoinactivated by ... More
Photoinactivation of fluorescein isothiocyanate-modified Na,K-ATPase by 2'(3')-O-(2,4,6-trinitrophenyl)8-azidoadenosine 5'-diphosphate. Abolition of E1 and E2 partial reactions by sequential block of high and low affinity nucleotide sites.
AuthorsWard DG, Cavieres JD
JournalJ Biol Chem
PubMed ID9603934
The Na,K-ATPase activity of the sodium pump exhibits apparent multisite kinetics toward ATP, a feature that is inherent to the minimal enzyme unit, the alpha beta protomer. We have argued that this should arise from separate catalytic and noncatalytic sites on the alpha beta protomer as fluorescein isothiocyanate (FITC) blocks ... More
Intrinsic fluorescence of the P-glycoprotein multidrug transporter: sensitivity of tryptophan residues to binding of drugs and nucleotides.
AuthorsLiu R, Siemiarczuk A, Sharom FJ
JournalBiochemistry
PubMed ID11101309
P-glycoprotein is a member of the ATP binding cassette family of membrane proteins, and acts as an ATP-driven efflux pump for a diverse group of hydrophobic drugs, natural products, and peptides. The side chains of aromatic amino acids have been proposed to play an important role in recognition and binding ... More
Interactions of Escherichia coli primary replicative helicase DnaB protein with nucleotide cofactors.
AuthorsJezewska MJ, Kim US, Bujalowski W
JournalBiophys J
PubMed ID8889182
Interactions between the Escherichia coli primary replicative helicase DnaB protein and nucleotide cofactors have been studied using several fluorescent nucleotide analogs and unmodified nucleotides. The thermodynamically rigorous fluorescent titration technique has been used to obtain true binding isotherms, independently of the assumptions of any relationships between the observed quenching of ... More
Inhibition of sodium and potassium adenosine triphosphatase by 2',3'-O-(2,4,6-trinitrocyclohexadienylidene) adenine nucleotides. Implications for the structure and mechanism of the Na:K pump.
AuthorsMoczydlowski EG, Fortes PA
JournalJ Biol Chem
PubMed ID6257716
Trinitrophenyl derivatives of adenine nucleotides (TNP-nucleotides: 2',3'-O-2,4,6-trinitrocyclohexadienylidene complexes at neutral or basic pH) are potent inhibitors of (Na,K)-ATPase activity. The inhibitory potency of the derivatives tested followed the sequence: TNP-ADP greater than TNP-ATP greater than TNP-AMP much greater than TNP-IMP greater than TNP-adenosine. In the presence of Na+ plus K+, ... More
Characterization of a phosphate binding domain on the alpha-subunit of chloroplast ATP synthase using the photoaffinity phosphate analogue 4-azido-2-nitrophenyl phosphate.
AuthorsGroth G, Mills DA, Christiansen E, Richter ML, Huchzermeyer B
JournalBiochemistry
PubMed ID11076517
The photoaffinity phosphate analogue 4-azido-2 nitrophenyl phosphate (ANPP) was shown previously (Pougeois, R., Lauquin, G. J.-M., and Vignais, P. V. (1983) Biochemistry 22, 1241-1245) to bind covalently and specifically to a single catalytic site on one of the three beta-subunits of the isolated chloroplast coupling factor 1 (CF(1)). Modification by ... More
Effect of nucleotides and actin on the orientation of the light chain-binding domain in myosin subfragment 1.
AuthorsSmyczynski C, Kasprzak AA
JournalBiochemistry
PubMed ID9341208
The X-ray structure of myosin head (S1) reveals the presence of a long alpha-helical structure that supports both the essential and the regulatory light chains. It has been proposed that small structural changes in the catalytic domain of S1 are amplified by swinging the long alpha-helix (the "lever arm") to ... More
An investigation of the SH1-SH2 and SH1-ATPase distances in myosin subfragment-1 by resonance energy transfer using nanosecond fluorimetry.
AuthorsCheung HC, Gonsoulin F, Garland F
JournalBiochim Biophys Acta
PubMed ID2932161
The separation between the two reactive thiols SH1 (Cys-704) and SH2 (Cys-694) and that between SH1 and the active site of myosin subfragment-1 were further investigated by Förster energy transfer techniques. The SH1-SH2 distance was determined with the probe 5-[[2-[(iodoacetyl)amino]ethyl] amino]naphthalene-1-sulfonic acid (AEDANS) attached to SH1 as the energy donor ... More
Resonance energy transfer between the adenosine 5'-diphosphate site of glutamate dehydrogenase and a guanosine 5'-triphosphate site containing a tyrosine labeled with 5'-[p-(fluorosulfonyl)benzoyl]-1,N6-ethenoadenosine.
AuthorsJacobson MA, Colman RF
JournalBiochemistry
PubMed ID6414507
The fluorescent nucleotide analogue 5'-[p-(fluorosulfonyl)benzoyl]-1,N6-ethenoadenosine (5'-FSB epsilon A) reacts irreversibly with bovine liver glutamate dehydrogenase and modifies one of the natural inhibitory guanosine 5'-triphosphate (GTP) sites [Jacobson, M.A., & Colman, R.F. (1982) Biochemistry 21, 2177-2186]. Enzyme with 1.28 mol of 5'-(p-sulfonylbenzoyl)-1,N6-ethenoadenosine/mol of subunit incorporated and exhibiting maximum change in sensitivity ... More
Negative cooperativity in the binding of nucleotides to Escherichia coli replicative helicase DnaB protein. Interactions with fluorescent nucleotide analogs.
AuthorsBujalowski W, Klonowska MM
JournalBiochemistry
PubMed ID8504109
The interactions of nucleotides with Escherichia coli replicative helicase DnaB protein have been systematically studied using fluorescent nucleotide analogs, 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP), 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-diphosphate (TNP-ADP), 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate (TNP-AMP), 3'-O-(N-methylantraniloyl) 5'-diphosphate (MANT-ADP), and 1,N6-ethenoadenosine diphosphate (epsilon ADP). The binding of the analogs is accompanied by strong quenching of the protein fluorescence; ... More
TNP-ATP and TNP-ADP as probes of the nucleotide binding site of CheA, the histidine protein kinase in the chemotaxis signal transduction pathway of Escherichia coli.
AuthorsStewart RC, VanBruggen R, Ellefson DD, Wolfe AJ
JournalBiochemistry
PubMed ID9724541
The interaction of CheA with ATP has important consequences in the chemotaxis signal transduction pathway of Escherichia coli. This interaction results in autophosphorylation of CheA, a histidine protein kinase. Autophosphorylation of CheA sets in motion a chain of biochemical events that enables the chemotaxis receptor proteins to communicate with the ... More
2',3'-O-(2,4,6-trinitrophenyl)-8-azido-adenosine mono-, di-, and triphosphates as photoaffinity probes of the Ca2+-ATPase of sarcoplasmic reticulum. Regulatory/superfluorescent nucleotides label the catalytic site with high efficiency.
AuthorsSeebregts CJ, McIntosh DB
JournalJ Biol Chem
PubMed ID2521624
We have synthesized a new class of ATP photo-affinity analogs, 2',3'-O-(2,4,6-trinitrophenyl)-8-azido (TNP-8N3)-ATP, -ADP, and -AMP, and their radiolabeled derivatives, and characterized their interaction with sarcoplasmic reticulum vesicles. The nucleotides bind with high affinity (Kd = 0.04-0.4 microM) to the catalytic site of the Ca2+-ATPase. TNP-8N3-ATP and TNP-8N3-ADP, at low concentrations ... More
Orientation of actin monomer in the F-actin filament: radial coordinate of glutamine-41 and effect of myosin subfragment 1 binding on the monomer orientation.
AuthorsKasprzak AA, Takashi R, Morales MF
JournalBiochemistry
PubMed ID3166995
We have employed the method of radial distance measurements in order to orient the actin monomer in the F-actin filament. This method utilizes fluorescence resonance energy transfer measurements of the distance between two equivalent chemical points located on two different monomers. The interprobe distance obtained this way is used to ... More
Cooperatively between catalytic sites in the mechanism of action of beef heart mitochondrial adenosine triphosphatase.
AuthorsGrubmeyer C, Penefsky HS
JournalJ Biol Chem
PubMed ID6452455
Occupancy of only one of two hydrolytic sites on beef heart mitochondrial ATPase (F1) by the radioactive ATP analog, 2',3'-O-(2,4,6-trinitrophenyl) adenosine 5'-[gamma-32P]-triphosphate (TNP-[gamma-32P]ATP) is associated with a low rate of hydrolysis of the substrate even under conditions otherwise favoring catalysis. Addition of excess nonradioactive TNP-ATP, in concentrations sufficient to fill ... More
Allosteric interactions between the nucleotide-binding sites and the ssDNA-binding site in the PriA helicase-ssDNA complex. 3.
AuthorsLucius AL, Jezewska MJ, Bujalowski W
JournalBiochemistry
PubMed ID16752913
Allosteric interactions between the strong and weak nucleotide-binding sites and the total and proper single-stranded (ss)DNA-binding sites of the Escherichia coli PriA helicase have been analyzed using the fluorescence titration technique. Binding of the DNA exclusively to the proper DNA-binding site of the helicase, profoundly affects the intrinsic affinities of ... More
Simultaneous binding of phosphate and TNP-ADP to FITC-modified NA+,K(+)-ATPase.
AuthorsScheiner-Bobis G, Antonipillai J, Farly RA
JournalBiochemistry
PubMed ID8396968
Double-reciprocal plots of the rate of ATP hydrolysis by Na+,K(+)-ATPase versus ATP concentration are not linear, and may reflect either two distinct binding sites for ATP or a single ATP binding site whose affinity for the nucleotide alternates between high-affinity and low-affinity states. In order to determine whether multiple nucleotides ... More
Nucleotide binding to the isolated beta subunit of the chloroplast ATP synthase.
AuthorsMills DA, Richter ML
JournalJ Biol Chem
PubMed ID1826906
The beta subunit isolated from the chloroplast ATP synthase F1 (CF1) has a single dissociable nucleotide binding site, consistent with the proposed function of this subunit in nucleotide binding and catalysis. The beta subunit bound the nucleotide analogs trinitrophenyl-ATP (TNP-ATP) or trinitrophenyl-ADP (TNP-ADP) with nearly equal affinities (Kd = 1-2 ... More
ATP regulation of sarcoplasmic reticulum Ca2+-ATPase. Metal-free ATP and 8-bromo-ATP bind with high affinity to the catalytic site of phosphorylated ATPase and accelerate dephosphorylation.
To localize and characterize the regulatory nucleotide site of skeletal muscle sarcoplasmic reticulum Ca2+-ATPase, we have investigated the effects of ADP, ATP, and analogues of these nucleotides on the rate of dephosphorylation of both native ATPase and ATPase modified with fluorescein 5'-isothiocyanate (FITC), a reagent which hinders access of nucleotides ... More
Characterization of a protease-resistant domain of the cytosolic portion of sarcoplasmic reticulum Ca2+-ATPase. Nucleotide- and metal-binding sites.
AuthorsChampeil P, Menguy T, Soulié S, Juul B, de Gracia AG, Rusconi F, Falson P, Denoroy L, Henao F, le Maire M, Moller JV
JournalJ Biol Chem
PubMed ID9506958
Treatment of rabbit sarcoplasmic reticulum Ca2+-ATPase with a variety of proteases, including elastase, proteinase K, and endoproteinases Asp-N and Glu-C, results in accumulation of soluble fragments starting close to the ATPase phosphorylation site Asp351 and ending in the Lys605-Arg615 region, well before the conserved sequences generally described as constituting the ... More
Characterization of the nucleotide binding properties of SV40 T antigen using fluorescent 3'(2')-O-(2,4,6-trinitrophenyl)adenine nucleotide analogues.
AuthorsHuang SG, Weisshart K, Fanning E
JournalBiochemistry
PubMed ID9799494
ATP binding to the large tumor (T) antigen encoded by the simian virus 40 (SV40) genome plays an essential role in the replication of viral DNA [Fanning, E., and Knippers, R. (1992) Annu. Rev. Biochem. 61, 55-85]. To better explore the functions of T antigen during the replication process, we ... More
Structural asymmetry of F1-ATPase caused by the gamma subunit generates a high affinity nucleotide binding site.
AuthorsKaibara C, Matsui T, Hisabori T, Yoshida M
JournalJ Biol Chem
PubMed ID8576203
The alpha 3 beta 3 gamma and alpha 3 beta 3 complexes of F1-ATPase from a thermophilic Bacillus PS3 were compared in terms of interaction with trinitrophenyl analogs of ATP and ADP (TNP-ATP and TNP-ADP) that differed from ATP and ADP and did not destabilize the alpha 3 beta 3 ... More
Inhibition by tentoxin of cooperativity among nucleotide binding sites on chloroplast coupling factor 1.
AuthorsHu N, Mills DA, Huchzermeyer B, Richter ML
JournalJ Biol Chem
PubMed ID8473298
Tentoxin, a cyclic tetrapeptide produced by the fungus Alternaria tenuis, is a potent inhibitor of the chloroplast coupling factor 1 from certain sensitive species of plants. We have shown that the beta subunit is at least partly responsible for conferring sensitivity to the toxin. This was confirmed by Avni et ... More
Distance relationships between the catalytic site labeled with 4-(iodoacetamido)salicylic acid and regulatory sites of glutamate dehydrogenase.
AuthorsJacobson MA, Colman RF
JournalBiochemistry
PubMed ID6487574
The distance between the catalytic site on bovine liver glutamate dehydrogenase labeled with 4-(iodoacetamido)salicylic acid (ISA) and the adenosine 5'-diphosphate (ADP) activatory site occupied by the analogue 2',3'-O-(2,4,6-trinitrocyclohexadienylidene)adenosine 5'-diphosphate (TNP-ADP) was evaluated by energy transfer. Native enzyme and enzyme containing about 1 mol of acetamidosalicylate/mol of subunit bind about 0.5 ... More
Transient kinetics of substrate binding to Na+/K(+)-ATPase measured by fluorescence quenching.
AuthorsPratap PR, Hellen EH, Palit A, Robinson JD
JournalBiophys Chem
PubMed ID9474752
This paper examines the transient kinetics of substrate binding to the Na+/K(+)-ATPase labelled with iodoacetamidofluorescein (IAF) using fluorescence quenching by trinitrophenyl-ATP (TNP-ATP). Earlier work (E.H. Hellen, P.R. Pratap, 1996, Fluorescence quenching of IAF-Na+/K(+)-ATPase via energy transfer to TNP-labelled nucleotide, Proceedings of the VIIIth International Conference on the Na+/K(+)-ATPase, in press) ... More