Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation
The 95 kDa subunit a of eukaryotic V-ATPases consists of a C-terminal, ion-translocating part and an N-terminal cytosolic domain. The latter’s N-terminal domain (~40 kDa) is described to bind in an acidification-dependent manner with cytohesin-2 (ARNO), giving the V-ATPase the putative function as p...
| Main Authors: | , , , , |
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| Format: | Journal Article |
| Language: | English |
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2013
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| Online Access: | https://hdl.handle.net/10356/104131 http://hdl.handle.net/10220/17044 |
| _version_ | 1826111258948009984 |
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| author | Dip, Phat Vinh Saw, Wuan Geok Roessle, Manfred Marshansky, Vladimir Grüber, Gerhard |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Dip, Phat Vinh Saw, Wuan Geok Roessle, Manfred Marshansky, Vladimir Grüber, Gerhard |
| author_sort | Dip, Phat Vinh |
| collection | NTU |
| description | The 95 kDa subunit a of eukaryotic V-ATPases consists of a C-terminal, ion-translocating part and an N-terminal cytosolic domain. The latter’s N-terminal domain (~40 kDa) is described to bind in an acidification-dependent manner with cytohesin-2 (ARNO), giving the V-ATPase the putative function as pH-sensing receptor. Recently, the solution structure of the very N-terminal segment of the cytosolic N-terminal domain has been solved. Here we produced the N-terminal truncated form SCa104-363 of the N-terminal domain (SCa1-363) of the Saccharomyces cerevisiae V-ATPase and determined its low resolution solution structure, derived from SAXS data. SCa104-363 shows an extended S-like conformation with a width of about 3.88 nm and a length of 11.4 nm. The structure has been superimposed into the 3D reconstruction of the related A1AO ATP synthase from Pyrococcus furiosus, revealing that the SCa104-363 fits well into the density of the collar structure of the enzyme complex. To understand the importance of the C-terminus of the protein SCa1-363, and to determine the localization of the N- and C-termini in SCa104-363, the C-terminal truncated form SCa106-324 was produced and analyzed by SAXS. Comparison of the SCa104-363 and SCa106-324 shapes showed that the additional loop region in SCa104-363 consists of the C-terminal residues. Whereas SCa104-363 is monomeric in solution, SCa106-324 forms a dimer, indicating the importance of the very C-terminus in structure formation. Finally, the solution structure of SCa104-363 and SCa106-324 will be discussed in terms of the topological arrangement of subunit a and cytoheisn-2 in V-ATPases. |
| first_indexed | 2024-10-01T02:47:45Z |
| format | Journal Article |
| id | ntu-10356/104131 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T02:47:45Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | ntu-10356/1041312020-03-07T12:24:55Z Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation Dip, Phat Vinh Saw, Wuan Geok Roessle, Manfred Marshansky, Vladimir Grüber, Gerhard School of Biological Sciences DRNTU::Science::Biological sciences The 95 kDa subunit a of eukaryotic V-ATPases consists of a C-terminal, ion-translocating part and an N-terminal cytosolic domain. The latter’s N-terminal domain (~40 kDa) is described to bind in an acidification-dependent manner with cytohesin-2 (ARNO), giving the V-ATPase the putative function as pH-sensing receptor. Recently, the solution structure of the very N-terminal segment of the cytosolic N-terminal domain has been solved. Here we produced the N-terminal truncated form SCa104-363 of the N-terminal domain (SCa1-363) of the Saccharomyces cerevisiae V-ATPase and determined its low resolution solution structure, derived from SAXS data. SCa104-363 shows an extended S-like conformation with a width of about 3.88 nm and a length of 11.4 nm. The structure has been superimposed into the 3D reconstruction of the related A1AO ATP synthase from Pyrococcus furiosus, revealing that the SCa104-363 fits well into the density of the collar structure of the enzyme complex. To understand the importance of the C-terminus of the protein SCa1-363, and to determine the localization of the N- and C-termini in SCa104-363, the C-terminal truncated form SCa106-324 was produced and analyzed by SAXS. Comparison of the SCa104-363 and SCa106-324 shapes showed that the additional loop region in SCa104-363 consists of the C-terminal residues. Whereas SCa104-363 is monomeric in solution, SCa106-324 forms a dimer, indicating the importance of the very C-terminus in structure formation. Finally, the solution structure of SCa104-363 and SCa106-324 will be discussed in terms of the topological arrangement of subunit a and cytoheisn-2 in V-ATPases. 2013-10-30T04:22:02Z 2019-12-06T21:27:08Z 2013-10-30T04:22:02Z 2019-12-06T21:27:08Z 2012 2012 Journal Article Dip, P. V., Saw, W. G., Roessle, M., Marshansky, V.,& Grüber, G. (2012). Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation. Journal of Bioenergetics and Biomembranes, 44(3), 341-350. 0145-479X https://hdl.handle.net/10356/104131 http://hdl.handle.net/10220/17044 10.1007/s10863-012-9442-3 en Journal of bioenergetics and biomembranes |
| spellingShingle | DRNTU::Science::Biological sciences Dip, Phat Vinh Saw, Wuan Geok Roessle, Manfred Marshansky, Vladimir Grüber, Gerhard Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation |
| title | Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation |
| title_full | Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation |
| title_fullStr | Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation |
| title_full_unstemmed | Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation |
| title_short | Solution structure of subunit a, a 104-363, of the Saccharomyces cerevisiae V-ATPase and the importance of its C-terminus in structure formation |
| title_sort | solution structure of subunit a a 104 363 of the saccharomyces cerevisiae v atpase and the importance of its c terminus in structure formation |
| topic | DRNTU::Science::Biological sciences |
| url | https://hdl.handle.net/10356/104131 http://hdl.handle.net/10220/17044 |
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