ATP hydrolysis is required to reset the ATP-binding cassette dimer into the resting-state conformation

被引:110
作者
Lu, G
Westbrooks, JM
Davidson, AL
Chen, J [1 ]
机构
[1] Purdue Univ, Dept Biol Sci, W Lafayette, IN 47907 USA
[2] Purdue Univ, Ctr Canc, W Lafayette, IN 47907 USA
[3] Baylor Coll Med, Dept Mol Virol & Microbiol, Houston, TX 77030 USA
关键词
ATP-binding cassette transporter; conformational change; posthydrolysis state; x-ray crystallography;
D O I
10.1073/pnas.0506039102
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
ATP-binding cassette (ABC) transporters couple ATP binding and hydrolysis to the movement of substances across the membrane; conformational changes clearly play an important role in the transporter mechanism. Previously, we have shown that a dimer of MaIK, the ATPase subunit of the maltose transporter from Escherichia coli, undergoes a tweezers-like motion in a transport cycle. The MaIK monomer consists of an N-terminal nucleotide binding domain and a C-terminal regulatory domain. The two nucleotide-binding domains in a dimer are either open or closed, depending on whether ATP is present, while the regulatory domains maintain contacts to hold the dinner together. In this work, the structure of MaIK in a posthydrolysis state is presented, obtained by cocrystallizing MalK with ATP-Mg2+. ATP was hydrolyzed in the crystallization drop, and ADP-Mg2+ was found in the resulting crystal structure. In contrast to the ATP-bound form where two ATP molecules are buried in a closed interface between the nucleotide-binding domains, the two nucleotide-binding domains of the ADP-bound form are open, indicating that ADP, unlike ATP, cannot stabilize the closed form. This conclusion is further supported by oligomerization studies of MaIK in solution. At low protein concentrations, ATP promotes dimerization of MalK, whereas ADP does not. The structures of dimeric MalK in the nucleotide-free, ATP-bound, and ADP-bound forms provide a framework for understanding the nature of the conformational changes that occur in an ATP-binding cassette transporter hydrolysis cycle, as well as how conformational changes in MaIK are coupled to solute transport.
引用
收藏
页码:17969 / 17974
页数:6
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