期刊
CELLULAR AND MOLECULAR LIFE SCIENCES
卷 67, 期 9, 页码 1519-1535出版社
SPRINGER BASEL AG
DOI: 10.1007/s00018-010-0266-1
关键词
VHH; Single-domain antibody; Gelsolin; Calcium; Immunomodulation; Cytoskeleton; Intrabody; Cancer cell
资金
- Fund for Scientific Research-Flanders (FWO-Vlaanderen)
- Stichting tegen Kanker
- Vlaamse Liga tegen Kanker
- Concerted Actions Program of Ghent University (GOA)
- Interuniversity attraction poles [IUAP06]
- VIB
- Vlaamse Liga tegen Kanker through a grant of the Stichting Emmanuel van der Schueren
RNA interference has tremendously advanced our understanding of gene function but recent reports have exposed undesirable side-effects. Recombinant Camelid single-domain antibodies (VHHs) provide an attractive means for studying protein function without affecting gene expression. We raised VHHs against gelsolin (GsnVHHs), a multifunctional actin-binding protein that controls cellular actin organization and migration. GsnVHH-induced delocalization of gelsolin to mitochondria or the nucleus in mammalian cells reveals distinct subpopulations including free gelsolin and actin-bound gelsolin complexes. GsnVHH 13 specifically recognizes Ca2+-activated gelsolin (K (d) similar to 10 nM) while GsnVHH 11 binds gelsolin irrespective of Ca2+ (K (d) similar to 5 nM) but completely blocks its interaction with G-actin. Both GsnVHHs trace gelsolin in membrane ruffles of EGF-stimulated MCF-7 cells and delay cell migration without affecting F-actin severing/capping or actin nucleation activities by gelsolin. We conclude that VHHs represent a potent way of blocking structural proteins and that actin nucleation by gelsolin is more complex than previously anticipated.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据