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Identification of a cysteine residue important for the ATPase activity of C. elegans fidgetin homologue
Author(s) -
Yakushiji Yasufumi,
Yamanaka Kunitoshi,
Ogura Teru
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.11.009
Subject(s) - cysteine , caenorhabditis elegans , atp hydrolysis , atpase , alanine , biochemistry , chemistry , residue (chemistry) , amino acid , biology , enzyme , microbiology and biotechnology , gene
Based on the amino acid alignment, Caenorhabditis elegans F32D1.1 was identified to be a homologue of the mammalian fidgetin. We produced and purified the F32D1.1 protein by using a baculovirus‐expression system. F32D1.1 has an ATPase activity, which is sensitive to N ‐ethylmaleimide. K m and V max for the ATPase activity of F32D1.1 were estimated to be 0.44 mM and 225 nmol/mg/min, respectively. When the cysteine at the position of 368 was mutated to alanine, the ATPase activity was greatly decreased; V max was decreased to one‐sixth, while K m remained similar. These results suggest that the unique position of cysteine 368, located immediately downstream of the Walker A motif, plays an important role in the ATP hydrolysis process of C. elegans F32D1.1 protein.