Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae
Proton-translocating vacuolar-type ATPases (V-ATPases) are necessary for numerous processes in eukaryotic cells, including receptor-mediated endocytosis, protein maturation, and lysosomal acidification. In mammals, V-ATPase subunit isoforms are differentially targeted to various intracellular compar...
| 主要な著者: | , , , , , |
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| フォーマット: | Journal article |
| 言語: | English |
| 出版事項: |
National Academy of Sciences
2019
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| _version_ | 1826294007444013056 |
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| author | Vasanthakumar, T Bueler, SA Wu, D Beilsten-Edmands, V Robinson, CV Rubinstein, JL |
| author_facet | Vasanthakumar, T Bueler, SA Wu, D Beilsten-Edmands, V Robinson, CV Rubinstein, JL |
| author_sort | Vasanthakumar, T |
| collection | OXFORD |
| description | Proton-translocating vacuolar-type ATPases (V-ATPases) are necessary for numerous processes in eukaryotic cells, including receptor-mediated endocytosis, protein maturation, and lysosomal acidification. In mammals, V-ATPase subunit isoforms are differentially targeted to various intracellular compartments or tissues, but how these subunit isoforms influence enzyme activity is not clear. In the yeast Saccharomyces cerevisiae, isoform diversity is limited to two different versions of the proton-translocating subunit a: Vph1p, which is targeted to the vacuole, and Stv1p, which is targeted to the Golgi apparatus and endosomes. We show that purified V-ATPase complexes containing Vph1p have higher ATPase activity than complexes containing Stv1p and that the relative difference in activity depends on the presence of lipids. We also show that VO complexes containing Stv1p could be readily purified without attached V1 regions. We used this effect to determine structures of the membrane-embedded VO region with Stv1p at 3.1-Å resolution, which we compare with a structure of the VO region with Vph1p that we determine to 3.2-Å resolution. These maps reveal differences in the surface charge near the cytoplasmic proton half-channel. Both maps also show the presence of bound lipids, as well as regularly spaced densities that may correspond to ergosterol or bound detergent, around the c-ring. |
| first_indexed | 2024-03-07T03:38:59Z |
| format | Journal article |
| id | oxford-uuid:bd3e5187-5830-4008-a462-30e2efb40cf9 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:38:59Z |
| publishDate | 2019 |
| publisher | National Academy of Sciences |
| record_format | dspace |
| spelling | oxford-uuid:bd3e5187-5830-4008-a462-30e2efb40cf92022-03-27T05:30:16ZStructural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiaeJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bd3e5187-5830-4008-a462-30e2efb40cf9EnglishSymplectic Elements at OxfordNational Academy of Sciences2019Vasanthakumar, TBueler, SAWu, DBeilsten-Edmands, VRobinson, CVRubinstein, JLProton-translocating vacuolar-type ATPases (V-ATPases) are necessary for numerous processes in eukaryotic cells, including receptor-mediated endocytosis, protein maturation, and lysosomal acidification. In mammals, V-ATPase subunit isoforms are differentially targeted to various intracellular compartments or tissues, but how these subunit isoforms influence enzyme activity is not clear. In the yeast Saccharomyces cerevisiae, isoform diversity is limited to two different versions of the proton-translocating subunit a: Vph1p, which is targeted to the vacuole, and Stv1p, which is targeted to the Golgi apparatus and endosomes. We show that purified V-ATPase complexes containing Vph1p have higher ATPase activity than complexes containing Stv1p and that the relative difference in activity depends on the presence of lipids. We also show that VO complexes containing Stv1p could be readily purified without attached V1 regions. We used this effect to determine structures of the membrane-embedded VO region with Stv1p at 3.1-Å resolution, which we compare with a structure of the VO region with Vph1p that we determine to 3.2-Å resolution. These maps reveal differences in the surface charge near the cytoplasmic proton half-channel. Both maps also show the presence of bound lipids, as well as regularly spaced densities that may correspond to ergosterol or bound detergent, around the c-ring. |
| spellingShingle | Vasanthakumar, T Bueler, SA Wu, D Beilsten-Edmands, V Robinson, CV Rubinstein, JL Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae |
| title | Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae |
| title_full | Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae |
| title_fullStr | Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae |
| title_full_unstemmed | Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae |
| title_short | Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae |
| title_sort | structural comparison of the vacuolar and golgi v atpases from saccharomyces cerevisiae |
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