Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits
Subunit G of the yeast V-ATPase is observed to be an elongated protein with a maximum size and the radius of gyration as 8.0 nm and 2.7 nm, respectively (ARMBRÜSTER et al., 2003). In order to understand the structure of subunit G and its possible orientation with respect to the other stalk subunits,...
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| Format: | Thesis |
| Language: | English |
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2012
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| Online Access: | https://hdl.handle.net/10356/47582 |
| _version_ | 1826130145203716096 |
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| author | Sankaranarayanan Rishikesan |
| author2 | Gerhard Gruber |
| author_facet | Gerhard Gruber Sankaranarayanan Rishikesan |
| author_sort | Sankaranarayanan Rishikesan |
| collection | NTU |
| description | Subunit G of the yeast V-ATPase is observed to be an elongated protein with a maximum size and the radius of gyration as 8.0 nm and 2.7 nm, respectively (ARMBRÜSTER et al., 2003). In order to understand the structure of subunit G and its possible orientation with respect to the other stalk subunits, two constructs G1-58 and G61-101, were created. The structure of both, the N- and C-terminal domains were solved by solution NMR technique. Precipitation assays have suggested the interaction of subunit G and E on the N-termini of both the subunits (JONES et al., 2005). Thus the interaction of subunit G with its neighboring stalk subunits, including E and d, were studied using NMR spectroscopy and FCS. Subunit E is an essential component of the V1 assembly, as disruption of the gene on the yeast (Vma4) resulted in a disassembly of the V1VO complex (KANE et al., 2003). The N-terminus of subunit E (E1-69) was expressed, purified to homogeneity and the solution structure was studied through NMR spectroscopy. The C-terminus of subunit E, a predicted globular domain E133-222 was cloned, expressed and purified to homogeneity. Efforts were also taken to study the solution structure of the same through NMR spectroscopy. The first secondary structure of E133-222 was determined and the region forming the β-strands on the globular domain is presented. |
| first_indexed | 2024-10-01T07:51:43Z |
| format | Thesis |
| id | ntu-10356/47582 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:51:43Z |
| publishDate | 2012 |
| record_format | dspace |
| spelling | ntu-10356/475822023-02-28T18:48:08Z Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits Sankaranarayanan Rishikesan Gerhard Gruber School of Biological Sciences DRNTU::Science::Biological sciences::Biochemistry Subunit G of the yeast V-ATPase is observed to be an elongated protein with a maximum size and the radius of gyration as 8.0 nm and 2.7 nm, respectively (ARMBRÜSTER et al., 2003). In order to understand the structure of subunit G and its possible orientation with respect to the other stalk subunits, two constructs G1-58 and G61-101, were created. The structure of both, the N- and C-terminal domains were solved by solution NMR technique. Precipitation assays have suggested the interaction of subunit G and E on the N-termini of both the subunits (JONES et al., 2005). Thus the interaction of subunit G with its neighboring stalk subunits, including E and d, were studied using NMR spectroscopy and FCS. Subunit E is an essential component of the V1 assembly, as disruption of the gene on the yeast (Vma4) resulted in a disassembly of the V1VO complex (KANE et al., 2003). The N-terminus of subunit E (E1-69) was expressed, purified to homogeneity and the solution structure was studied through NMR spectroscopy. The C-terminus of subunit E, a predicted globular domain E133-222 was cloned, expressed and purified to homogeneity. Efforts were also taken to study the solution structure of the same through NMR spectroscopy. The first secondary structure of E133-222 was determined and the region forming the β-strands on the globular domain is presented. DOCTOR OF PHILOSOPHY (SBS) 2012-01-09T07:06:19Z 2012-01-09T07:06:19Z 2011 2011 Thesis Sankaranarayanan Rishikesan. (2011). Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits. Doctoral thesis, Nanyang Technological University, Singapore. https://hdl.handle.net/10356/47582 10.32657/10356/47582 en 138 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences::Biochemistry Sankaranarayanan Rishikesan Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits |
| title | Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits |
| title_full | Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits |
| title_fullStr | Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits |
| title_full_unstemmed | Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits |
| title_short | Solution structure of the stalk subunits E and G of the vacuolar ATPase from yeast and interaction studies of subunits E and G with the neighboring subunits |
| title_sort | solution structure of the stalk subunits e and g of the vacuolar atpase from yeast and interaction studies of subunits e and g with the neighboring subunits |
| topic | DRNTU::Science::Biological sciences::Biochemistry |
| url | https://hdl.handle.net/10356/47582 |
| work_keys_str_mv | AT sankaranarayananrishikesan solutionstructureofthestalksubunitseandgofthevacuolaratpasefromyeastandinteractionstudiesofsubunitseandgwiththeneighboringsubunits |