| Ligand-dependent structural changes in the V(1) ATPase from Manduca sexta. | |
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MedLine Citation:
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PMID: 15337855 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The response of V(1) ATPase of the tobacco hornworm Manduca sexta to Mg(2+) and nucleotide binding in the presence of the enhancer methanol has been studied by CuCl(2)-induced disulfide formation, fluorescence spectroscopy, and small-angle X-ray scattering. When the V(1) complex was supplemented with CuCl(2) nucleotide-dependence of A-B-E and A-B-E-D cross-linking products was observed in absence of nucleotides and presence of MgADP+Pi but not when MgAMP.PNP or MgADP were added. A zero-length cross-linking product of subunits D and E was formed, supporting their close proximity in the V(1) complex. The catalytic subunit A was reacted with N-4[4-[7-(dimethylamino)-4-methyl]coumarin-3-yl]maleimide (CM) and spectral shifts and changes in fluorescence intensity were detected upon addition of MgAMP.PNP, -ATP, -ADP+Pi, or -ADP. Differences in the fluorescence emission of these nucleotide-binding states were monitored using the intrinsic tryptophan fluorescence. The structural composition of the V(1) ATPase from M. sexta and conformational alterations in this enzyme due to Mg(2+) and nucleotide binding are discussed on the basis of these and previous observations. |
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Authors:
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Unal Coskun; Vincenzo F Rizzo; Michel H J Koch; Gerhard Grüber |
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Publication Detail:
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Type: Comparative Study; Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of bioenergetics and biomembranes Volume: 36 ISSN: 0145-479X ISO Abbreviation: J. Bioenerg. Biomembr. Publication Date: 2004 Jun |
Date Detail:
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Created Date: 2004-08-31 Completed Date: 2005-03-07 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 7701859 Medline TA: J Bioenerg Biomembr Country: United States |
Other Details:
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Languages: eng Pagination: 249-56 Citation Subset: IM |
Affiliation:
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Universität des Saarlandes, Fachrichtung 2.5 - Biophysik, D-66421 Homburg, Germany. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Copper Cross-Linking Reagents / metabolism Ligands Magnesium / pharmacology Manduca / enzymology* Methanol / metabolism Nucleotides / metabolism Protein Conformation / drug effects Scattering, Radiation Spectrometry, Fluorescence Vacuolar Proton-Translocating ATPases / isolation & purification*, metabolism* X-Rays |
| Chemical | |
Reg. No./Substance:
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0/Cross-Linking Reagents; 0/Ligands; 0/Nucleotides; 67-56-1/Methanol; 7439-95-4/Magnesium; 7440-50-8/Copper; 7447-39-4/cupric chloride; EC 3.6.1.-/Vacuolar Proton-Translocating ATPases |
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