A Different Conformation for EGC Stator Subcomplex in Solution and in the Assembled Yeast V-ATPase: Possible Implications for Regulatory Disassembly

被引:55
作者
Diepholz, Meikel [1 ]
Venzke, David [1 ]
Prinz, Simone [1 ]
Batisse, Claire [1 ]
Floerchinger, Beate [1 ]
Roessle, Manfred [2 ]
Svergun, Dmitri I. [2 ,3 ]
Boettcher, Bettina [1 ,4 ]
Fethiere, James [5 ,6 ]
机构
[1] EMBL, Struct & Computat Biol Unit, D-69117 Heidelberg, Germany
[2] EMBL, Hamburg Outstn, D-22603 Hamburg, Germany
[3] Russian Acad Sci, Inst Crystallog, Moscow 117333, Russia
[4] Univ Edinburgh, Sch Biol Sci, Edinburgh EH9 3JR, Midlothian, Scotland
[5] Univ Montreal, Inst Rech Immunol & Cancerol, Montreal, PQ H3C 3J7, Canada
[6] Univ Montreal, Dept Pharmacol, Montreal, PQ H3C 3J7, Canada
基金
美国国家卫生研究院;
关键词
D O I
10.1016/j.str.2008.09.010
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Vacuolar ATPases (V-ATPases) are ATP-dependent proton pumps that maintain the acidity of cellular compartments. They are composed of a membrane-integrated proton-translocating V-0 and an extrinsic cytoplasmic catalytic domain V-1, joined by several connecting subunits. To clarify the arrangement of these peripheral connections and their interrelation with other subunits of the holocomplex, we have determined the solution structures of isolated EG and EGC connecting subcomplexes by small angle X-ray scattering and the 3D map of the yeast V-ATPase by electron microscopy. In solution, EG forms a slightly kinked rod, which assembles with subunit C into an L-shaped structure. This model is supported by the microscopy data, which show three copies of EG with two of these linked by subunit C. However, the relative arrangement of the EG and C subunits in solution is more open than that in the holoenzyme, suggesting a conformational change of EGC during regulatory assembly and disassembly.
引用
收藏
页码:1789 / 1798
页数:10
相关论文
共 51 条
[1]   Cysteine-directed cross-linking to subunit B suggests that subunit E forms part of the peripheral stalk of the vacuolar H+-ATPase [J].
Arata, Y ;
Baleja, JD ;
Forgac, M .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (05) :3357-3363
[2]   Structural analysis of the stalk subunit Vma5p of the yeast V-ATPase in solution [J].
Armbrüster, A ;
Svergun, DI ;
Coskun, U ;
Juliano, S ;
Bailer, SM ;
Grüber, G .
FEBS LETTERS, 2004, 570 (1-3) :119-125
[3]   Three-dimensional structure of the intact Thermus thermophilus H+-ATPase/synthase by electron microscopy [J].
Bernal, RA ;
Stock, D .
STRUCTURE, 2004, 12 (10) :1789-1798
[4]   Biological motors - Connecting stalks in V-type ATPase [J].
Boekema, EJ ;
van Breemen, JFL ;
Brisson, A ;
Ubbink-Kok, T ;
Konings, WN ;
Lolkema, JS .
NATURE, 1999, 401 (6748) :37-38
[5]   Crystal structure of yeast V-ATPase subunit C reveals its stator function [J].
Drory, O ;
Frolow, F ;
Nelson, N .
EMBO REPORTS, 2004, 5 (12) :1148-1152
[6]   Stoichiometry and localization of the stator subunits E and G in Thermus thermophilus H+-ATPase/synthase [J].
Esteban, Olga ;
Bernal, Ricardo A. ;
Donohoe, Mhairi ;
Videler, Hortense ;
Sharon, Michal ;
Robinson, Carol V. ;
Stock, Daniela .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2008, 283 (05) :2595-2603
[7]   Peripheral stator of the yeast V-ATPase:: Stoichiometry and specificity of interaction between the EG complex and subunits C and H [J].
Féthière, J ;
Venzke, D ;
Madden, DR ;
Böttcher, B .
BIOCHEMISTRY, 2005, 44 (48) :15906-15914
[8]   Building the stator of the yeast vacuolar-ATPase -: Specific interaction between subunits E and G [J].
Féthière, J ;
Venzke, D ;
Diepholz, M ;
Seybert, A ;
Geerlof, A ;
Gentzel, M ;
Wilm, M ;
Böttcher, B .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (39) :40670-40676
[9]   Vacuolar ATPases: rotary proton pumps in physiology and pathophysiology [J].
Forgac, Michoel .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2007, 8 (11) :917-929
[10]   SPIDER and WEB: Processing and visualization of images in 3D electron microscopy and related fields [J].
Frank, J ;
Radermacher, M ;
Penczek, P ;
Zhu, J ;
Li, YH ;
Ladjadj, M ;
Leith, A .
JOURNAL OF STRUCTURAL BIOLOGY, 1996, 116 (01) :190-199