| Characterization of exchangeable and nonexchangeable bound adenine nucleotides in F1-ATPase from Escherichia coli. | |
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MedLine Citation:
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PMID: 2867094 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The total amount of bound exchangeable and nonexchangeable adenine nucleotides in Escherichia coli F1-ATPase (BF1) was determined; three exchangeable nucleotides were assessed by equilibrium dialysis in a [14C]ADP-supplemented medium. When BF1 was purified in a medium supplemented with ATP, a stoichiometry of nearly 6 mol of bound nucleotides/mol of enzyme was found; three of the bound nucleotides were ATP and the others ADP. When BF1 was filtered on Sephadex G-50 in a glycerol medium (Garrett, N.E., and Penefsky, H.S. (1975) J. Biol. Chem. 250, 6640-6647), bound ADP was rapidly released, in contrast to bound ATP which remained firmly attached to the enzyme. Upon incubation of BF1 with [14C]ADP, the bound ADP rather than the bound ATP was exchanged. Of the three [14C]ADPs which have bound to BF1 by exchange after equilibrium dialysis, one was readily lost by gel filtration on Sephadex G-50; the loss of bound [14C]ADP was markedly reduced by incubation of BF1 with aurovertin, a specific ligand of the beta subunit which is known to increase the affinity of the beta subunit for nucleotides (Issartel, J.-P., and Vignais, P. V. (1984) Biochemistry 23, 6591-6595). Upon photoirradiation of BF1 with [alpha-32P]2-azido-ADP, only the beta subunit was labeled; concomitantly, bound ADP was released, but the content in bound ATP remained stable. These results suggest that specific sites located on the three beta subunits bind nucleotides in a reversible manner. Consequently, the tightly bound ATP of native BF1 would be located on the alpha subunits. |
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Authors:
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J P Issartel; J Lunardi; P V Vignais |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The Journal of biological chemistry Volume: 261 ISSN: 0021-9258 ISO Abbreviation: J. Biol. Chem. Publication Date: 1986 Jan |
Date Detail:
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Created Date: 1986-02-14 Completed Date: 1986-02-14 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 2985121R Medline TA: J Biol Chem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 895-901 Citation Subset: IM |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adenine Nucleotides
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metabolism* Adenosine Diphosphate / analogs & derivatives, metabolism Adenosine Triphosphate / metabolism Aurovertins / pharmacology Azides* Dose-Response Relationship, Drug Escherichia coli / enzymology* Kinetics Macromolecular Substances Proton-Translocating ATPases / metabolism* Time Factors |
| Chemical | |
Reg. No./Substance:
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0/Adenine Nucleotides; 0/Aurovertins; 0/Azides; 0/Macromolecular Substances; 56-65-5/Adenosine Triphosphate; 58-64-0/Adenosine Diphosphate; 64020-53-7/2-azidoadenosine 3',5'-diphosphate; EC 3.6.3.14/Proton-Translocating ATPases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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