Detection of calmodulin-binding proteins and calmodulin-dependent phosphorylation linked to calmodulin-dependent chemotaxis to folic and cAMP in Dictyostelium

被引:30
作者
Gauthier, ML [1 ]
O'Day, DH [1 ]
机构
[1] Univ Toronto, Dept Zool, Mississauga, ON L5L 1C6, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
calmodulin; CaM-binding proteins; chemotaxis; protein phosphorylation; Dictyostelium;
D O I
10.1016/S0898-6568(01)00187-5
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Calmodulin (CaM) antagonists, trifluoperazine (TFP) or calmidazolium (R24571), dose-dependently inhibited cAMP and folic acid (FA) chemotaxis in Dictyostelium. Developing, starved, and refed cells were compared to determine if certain CaM-binding proteins (CaMBPs) and CaM-dependent phosphorylation events could be identified as potential downstream effectors. Recombinant CaM ([S-35]VU-1-CaM) gel overlays coupled with cell fractionation revealed at least three dozen Ca2+-dependent and around 12 Ca2+-independent CaMBPs in Dictyostelium. The CaMBPs associated with early development were also found in experimentally starved cells (cAMP chemotaxis), but were different for the CaMBP population linked to growth-phase cells (FA chemotaxis). Probing Western blots with phosphoserine antibodies revealed several phosphoprotein bands that displayed increases when cAMP-responsive cells were treated with TFP. In FA-responsive cells, several but distinct phosphoproteins decreased when treated with TFP. These data show that unique CaMBPs are present in growing, FA-chemosensitive cells vs. starved cAMP-chemoresponsive cells that may be important for mediating CaM-dependent events during chemotaxis. (C) 2001 Elsevier Science Inc. All rights reserved.
引用
收藏
页码:575 / 584
页数:10
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