Identification of a cysteine residue important for the ATPase activity of C. elegans fidgetin homologue
Identification of a cysteine residue important for the ATPase activity of C. elegans fidgetin homologue
复制标题
DOI：
10.1016/j.febslet.2004.11.009
复制
发表时间：
2004-12
期刊：
影响因子：
3.5
通讯作者：
Y. Yakushiji;K. Yamanaka;T. Ogura
中科院分区：
文献类型：
--
作者：
Y. Yakushiji;K. Yamanaka;T. Ogura
基于氨基酸比对，秀丽隐杆线虫F32D1.1被鉴定为哺乳动物fidgetin的同源物。我们通过杆状病毒表达系统生产并纯化了F32D1.1蛋白。F32D1.1具有ATP酶活性，对N-乙基马来酰亚胺敏感。F_（32）D_（1.1）ATP酶活性的Km和Vmax分别为0.44 mM和225 nmol/mg/min。当368位的半胱氨酸突变为丙氨酸时，ATP酶活性大大降低，Vmax降低到六分之一，而Km保持不变。这些结果表明，位于步行者A基序下游的368位半胱氨酸在C. elegans F32D1.1蛋白。
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. Kmand Vmaxfor 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; Vmaxwas decreased to one-sixth, while Kmremained 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.