WAVE Regulatory Complex Activation

被引：10

作者：

Hume, Peter J. ^{[1
]}

Humphreys, Daniel ^{[1
]}

Koronakis, Vassilis ^{[1
]}

机构：

[1] Univ Cambridge, Dept Pathol, Cambridge CB2 1QP, England

来源：

RECONSTITUTING THE CYTOSKELETON | 2014年 / 540卷

基金：

英国惠康基金;

关键词：

ACTIN; SALMONELLA; LIPOSOMES; MEMBRANE; ARF6; WASP;

D O I：

10.1016/B978-0-12-397924-7.00020-0

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The WAVE regulatory complex (WRQ is critical to control of actin polymerization at the eukaryotic cell membrane. By reconstituting WAVE-dependent actin assembly on silica microspheres coated with phospholipid bilayers in mammalian brain extracts, we discovered that membrane recruitment and activation of WRC require the cooperative action of two mammalian GTPases, Aft and Rac. Here, we describe detailed methods to generate phospholipid-coated microspheres and porcine brain extract and outline conditions necessary to reconstitute WRC-dependent motility. In addition, we describe how to generate acylated recombinant GTPases, anchor them to lipid-coated microspheres, and reconstitute GTPase activation of WRC.

引用

页码：363 / 379

页数：17

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