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磷脂酶C-δ1的D-肌醇1,4,5-三磷酸结合结构域

D-myo-inositol 1,4,5-trisphosphate binding domain of phospholipase C-delta 1.

作者信息

Hirata M, Kanematsu T, Sakuma K, Koga T, Watanabe Y, Ozaki S, Yagisawa H

机构信息

Department of Biochemistry, Faculty of Dentistry, Kyushu University, Fukuoka, Japan.

出版信息

Biochem Biophys Res Commun. 1994 Dec 30;205(3):1563-71. doi: 10.1006/bbrc.1994.2845.

Abstract

D-myo-Inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) binding domain of phospholipase C-delta 1 (PLC-delta 1) was determined by examining binding activity of the synthetic peptide corresponding to residues 30-43 of PLC-delta 1. The peptide coupled with carrier proteins such as keyhole limpet hemocyanin or bovine serum albumin bound Ins(1,4,5)P3, whereas a scrambled peptide with the same amino acids did not do so. Polyclonal antibody against the peptide was examined to determine whether it would cause inhibition of the Ins(1,4,5)P3 binding to PLC-delta 1. Fab fragment of antibody to the peptide did inhibit binding to PLC-delta 1, in a dose-dependent manner. Thus it seems likely that the region of residues 30-43 of PLC-delta 1 is responsible for the binding of Ins(1,4,5)P3.

摘要

通过检测对应于磷脂酶C-δ1(PLC-δ1)第30 - 43位残基的合成肽的结合活性,确定了磷脂酶C-δ1(PLC-δ1)的D-肌醇1,4,5-三磷酸(Ins(1,4,5)P3)结合结构域。与诸如钥孔血蓝蛋白或牛血清白蛋白等载体蛋白偶联的该肽能结合Ins(1,4,5)P3,而具有相同氨基酸的乱序肽则不能。检测了针对该肽的多克隆抗体,以确定其是否会抑制Ins(1,4,5)P3与PLC-δ1的结合。该肽的抗体Fab片段确实以剂量依赖性方式抑制了与PLC-δ1的结合。因此,PLC-δ1的第30 - 43位残基区域似乎负责Ins(1,4,5)P3的结合。

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