par Van Den Abbeele, Anske;De Clercq, Sarah 
;De Ganck, Ariane;De Corte, Veerle;Van Loo, Berlinda;Soror, Sameh Hamdy;Srinivasan, Vasundara;Steyaert, Jan;Vandekerckhove, Joël;Gettemans, Jan
Référence Cellular and molecular life sciences, 67, 9, page (1519-1535)
Publication Publié, 2010-05
;De Ganck, Ariane;De Corte, Veerle;Van Loo, Berlinda;Soror, Sameh Hamdy;Srinivasan, Vasundara;Steyaert, Jan;Vandekerckhove, Joël;Gettemans, JanRéférence Cellular and molecular life sciences, 67, 9, page (1519-1535)
Publication Publié, 2010-05
Article révisé par les pairs
| Résumé : | 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 ~10 nM) while GsnVHH 11 binds gelsolin irrespective of Ca 2+ (K d ~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. © 2010 Birkhäuser Verlag. |
