DELETION AND SITE-DIRECTED MUTAGENESIS OF EF-HAND DOMAIN OF PHOSPHOLIPASE C-DELTA-1 - EFFECTS ON ITS ACTIVITY

被引：37

作者：

NAKASHIMA, S

BANNO, Y

WATANABE, T

NAKAMURA, Y

MIZUTANI, T

SAKAI, H

ZHAO, YT

SUGIMOTO, Y

NOZAWA, Y

机构：

[1] GIFU UNIV,SCH MED,DEPT BIOCHEM,GIFU 500,JAPAN

[2] SHIRAKAWA INST ANIM GENET,NISHI SHIRAKAWA,FUKUSHIMA 961,JAPAN

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1995年 / 211卷 / 02期

关键词：

D O I：

10.1006/bbrc.1995.1822

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In order to elucidate a role of a putative EF-hand motif (144-172) in phospholipase C-delta 1 (PLC-FI), deletion and point mutation of the enzyme were performed and the mutated cDNAs were expressed in CHO cells and E. coli AD202 strain. Deletion of amino acid residues of 141-236 or 173-236 resulted in abolition of PLC activity. However, the decreased PLC activity to 15-20% by deletion of the EF-hand motif (144-172) was still Ca2+-dependent. Furthermore, mutants, in which conserved Asp(153), Asp(157), Glu(164) or all these acidic amino acids in the EF-hand motif were replaced with alanine residues, showed nearly the same PLC activity and Ca2+-dependency as those of wild-type. These results suggest that the region containing the EF-hand motif may not play a role in regulation of Ca2+-sensitivity of PLC-delta 1, but is important for its activity. (C) 1995 Academic Press, Inc.

引用

页码：364 / 369

页数：6

共 21 条

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