Inhibition of F1-ATPase Rotational Catalysis by the Carboxyl-terminal Domain of the ε Subunit

被引:13
作者
Nakanishi-Matsui, Mayumi [1 ]
Sekiya, Mizuki [1 ]
Yano, Shio [1 ]
Futai, Masamitsu [1 ]
机构
[1] Iwate Med Univ, Fac Pharmaceut Sci, Dept Biochem, Yahaba, Iwate 0283694, Japan
基金
日本学术振兴会;
关键词
COLI ATP SYNTHASE; ESCHERICHIA-COLI; F-1; SECTOR; GAMMA-SUBUNIT; MECHANISM; COMPLEX; BINDING; FOF1;
D O I
10.1074/jbc.M114.578872
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
Escherichia coli ATP synthase (F0F1) couples catalysis and proton transport through subunit rotation. The epsilon subunit, an endogenous inhibitor, lowers F-1-ATPase activity by decreasing the rotation speed and extending the duration of the inhibited state (Sekiya, M., Hosokawa, H., Nakanishi-Matsui, M., Al-Shawi, M. K., Nakamoto, R. K., and Futai, M. (2010) Single molecule behavior of inhibited and active states of Escherichia coli ATP synthase F-1 rotation. J. Biol. Chem. 285, 42058-42067). In this study, we constructed a series of epsilon subunits truncated successively from the carboxyl-terminal domain (helix 1/loop 2/helix 2) and examined their effects on rotational catalysis (ATPase activity, average rotation rate, and duration of inhibited state). As expected, the epsilon subunit lacking helix 2 caused about 1/2-fold reduced inhibition, and that without loop 2/helix 2 or helix 1/loop 2/helix 2 showed a further reduced effect. Substitution of epsilon Ser(108) in loop 2 and epsilon Tyr(114) in helix 2, which possibly interact with the beta and gamma subunits, respectively, decreased the inhibitory effect. These results suggest that the carboxyl-terminal domain of the epsilon subunit plays a pivotal role in the inhibition of F-1 rotation through interaction with other subunits.
引用
收藏
页码:30822 / 30831
页数:10
相关论文
共 36 条
[1]   STRUCTURE AT 2.8-ANGSTROM RESOLUTION OF F1-ATPASE FROM BOVINE HEART-MITOCHONDRIA [J].
ABRAHAMS, JP ;
LESLIE, AGW ;
LUTTER, R ;
WALKER, JE .
NATURE, 1994, 370 (6491) :621-628
[2]  
AlShawi MK, 1997, J BIOL CHEM, V272, P2300
[3]   Conformation of the γ subunit at the γ-ε-c interface in the complete Escherichia coli F1-ATPase complex by site-directed spin labeling [J].
Andrews, SH ;
Peskova, YB ;
Polar, MK ;
Herlihy, VB ;
Nakamoto, RK .
BIOCHEMISTRY, 2001, 40 (35) :10664-10670
[4]   High-resolution single-molecule characterization of the enzymatic states in Escherichia coli F1-ATPase [J].
Bilyard, Thomas ;
Nakanishi-Matsui, Mayumi ;
Steel, Bradley C. ;
Pilizota, Teuta ;
Nord, Ashley L. ;
Hosokawa, Hiroyuki ;
Futai, Masamitsu ;
Berry, Richard M. .
PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES, 2013, 368 (1611)
[5]   Spotlighting motors and controls of single FoF1-ATP synthase [J].
Boersch, Michael ;
Duncan, Thomas M. .
BIOCHEMICAL SOCIETY TRANSACTIONS, 2013, 41 :1219-1226
[6]   The ATP synthase - A splendid molecular machine [J].
Boyer, PD .
ANNUAL REVIEW OF BIOCHEMISTRY, 1997, 66 :717-749
[7]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[8]   Rotor/stator interactions of the ε subunit in Escherichia coli ATP synthase and implications for enzyme regulation [J].
Bulygin, VV ;
Duncan, TM ;
Cross, RL .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (34) :35616-35621
[9]   Structure of the ATP synthase catalytic complex (F1) from Escherichia coli in an autoinhibited conformation [J].
Cingolani, Gino ;
Duncan, Thomas M. .
NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2011, 18 (06) :701-U100
[10]   Genetic fusions of globular proteins to the ε subunit of the Escherichia coli ATP synthase -: Implications for in vivo rotational catalysis and ε subunit function [J].
Cipriano, DJ ;
Bi, YM ;
Dunn, SD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (19) :16782-16790