Three-dimensional structure and subunit topology of the V-1 ATPase from Manduca sexta midgut

DC FieldValueLanguage
dc.contributor.authorGruber, G
dc.contributor.authorRadermacher, M
dc.contributor.authorRuiz, T
dc.contributor.authorGodovac-Zimmermann, J
dc.contributor.authorCanas, B
dc.contributor.authorKleine-Kohlbrecher, D
dc.contributor.authorHuss, M
dc.contributor.authorHarvey, WR
dc.contributor.authorWieczorek, H
dc.date.accessioned2021-12-23T16:21:29Z-
dc.date.available2021-12-23T16:21:29Z-
dc.date.issued2000
dc.identifier.issn00062960
dc.identifier.urihttps://osnascholar.ub.uni-osnabrueck.de/handle/unios/13892-
dc.description.abstractThe three-dimensional structure of the Manduca sexta midgut V-1 ATPase has been determined at 3.2 nm resolution from electron micrographs of negatively stained specimens. The V-1 complex has a barrel-like structure Il nm in height and 13.5 nm in diameter. It is hexagonal in the top view, whereas in the side view, the six large subunits A and B are interdigitated for most of their length (9 nm). The topology and importance of the individual subunits of the V-1 complex have been explored by protease digestion, resistance to chaotropic agents, MALDI-TOF mass spectrometry, and CuCl2-induced disulfide formation. Treatment of V-1 with trypsin or chaotropic iodide resulted in a rapid cleavage or release of subunit D from the enzyme, indicating that this subunit is exposed in the complex. Trypsin cleavage of V-1 decreased the ATPase activity with a time course that was in line with the cleavage of subunits B, C, G, and F. When CuCl2 was added to V-1 in the presence of CaADP, the cross-linked products A-E-F and B-H were generated. In experiments where CuCl2 was added after preincubation of CaATP, the cross-linked products E-F and E-G were formed. These changes in cross-linking of subunit E to near-neighbor subunits support the hypothesis that these are nucleotide-dependent conformational changes of the E subunit.
dc.description.sponsorshipPHS HHSUnited States Department of Health & Human ServicesUnited States Public Health Service [A1 22444] Funding Source: Medline
dc.language.isoen
dc.publisherAMER CHEMICAL SOC
dc.relation.ispartofBIOCHEMISTRY
dc.subject3-DIMENSIONAL RECONSTRUCTION
dc.subjectBiochemistry & Molecular Biology
dc.subjectCRYOELECTRON MICROSCOPY
dc.subjectELECTRON-MICROSCOPY
dc.subjectESCHERICHIA-COLI
dc.subjectF1 ADENOSINE-TRIPHOSPHATASE
dc.subjectGAMMA-SUBUNIT
dc.subjectMOLECULAR ARCHITECTURE
dc.subjectTOBACCO HORNWORM MIDGUT
dc.subjectV-TYPE ATPASE
dc.subjectVACUOLAR-TYPE ATPASE
dc.titleThree-dimensional structure and subunit topology of the V-1 ATPase from Manduca sexta midgut
dc.typejournal article
dc.identifier.doi10.1021/bi000103u
dc.identifier.isiISI:000088376900029
dc.description.volume39
dc.description.issue29
dc.description.startpage8609
dc.description.endpage8616
dc.contributor.researcheridE-6863-2010
dc.contributor.researcheridO-9256-2019
dc.publisher.place1155 16TH ST, NW, WASHINGTON, DC 20036 USA
dcterms.isPartOf.abbreviationBiochemistry
crisitem.author.deptUniversität Osnabrück-
crisitem.author.deptFB 05 - Biologie/Chemie-
crisitem.author.deptidfb05-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.netidHuMa001-
crisitem.author.netidWiHe990-
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