Mutational analysis of the functional motifs in the ATPase domain of Caenorhabditis elegans fidgetin homologue FIGL-1: firm evidence for an intersubunit catalysis mechanism of ATP hydrolysis by AAA ATPases

Journal of Structural Biology
Yasufumi YakushijiTeru Ogura

Abstract

The AAA family proteins usually form a hexameric ring structure. The ATP-binding pocket, which is located at the interface of subunits in the hexamer, consists of three functionally important motifs, the Walker A and B motifs, and the second region of homology (SRH). It is well known that Walker A and B motifs mediate ATP binding and hydrolysis, respectively. Highly conserved arginine residues in the SRH have been proposed to function as arginine fingers, which interact with the gamma-phosphate of bound ATP. To elucidate the mechanism of ATP hydrolysis, we prepared several mutants of the Caenorhabditis elegans fidgetin homologue FIGL-1 carrying a mutation in each of the above-mentioned three motifs. None of the constructed mutants showed ATPase activity. All the mutants except for K362A were able to bind ATP. A decrease in the ATPase activity by mixing wild-type and each mutant subunits was caused by the formation of hetero-hexamers. Mixtures of E416A and R471A, or N461A and R471A led to the formation of hetero-hexamers with partially restored ATPase activities, providing direct, firm evidence for the intersubunit catalysis model. In addition, based on the results obtained with mixtures of K362A with wild-type or R471A subunits...Continue Reading

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Jul 28, 2001·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·T Ogura, A J Wilkinson
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Citations

Jul 22, 2009·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Yuka Matsushita-IshiodoriTeru Ogura
Apr 14, 2009·Journal of Molecular Biology·Jingting WangSuzanne Scarlata
Feb 28, 2013·Biophysical Journal·Timothy TraversCarlos J Camacho
Oct 11, 2015·Biochimica Et Biophysica Acta·Immanuel GrimmWolfgang Girzalsky
Feb 5, 2010·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Masatoshi Esaki, Teru Ogura
Mar 15, 2018·The Journal of Cell Biology·Coralie FassierJamilé Hazan
Jan 17, 2021·Molecular & Cellular Proteomics : MCP·Yadong YuYifan Cheng
May 12, 2012·Journal of Structural Biology·Akinobu OnitakeTeru Ogura

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