Title | A llama-derived gelsolin single-domain antibody blocks gelsolin-G-actin interaction. |
Publication Type | Journal Article |
Year of Publication | 2010 |
Authors | Van den Abbeele, A., S. De Clercq, A. De Ganck, V. De Corte, B. Van Loo, S. Hamdy Soror, V. Srinivasan, J. Steyaert, J. Vandekerckhove, and J. Gettemans |
Journal | Cell Mol Life Sci |
Volume | 67 |
Issue | 9 |
Pagination | 1519-35 |
Date Published | 2010 May |
ISSN | 1420-9071 |
Keywords | Actins, Animals, Calcium, Camelids, New World, Cell Line, Cell Movement, Crystallography, X-Ray, Epitopes, Gelsolin, Humans, Mitochondria, Models, Molecular, Molecular Sequence Data, Protein Structure, Tertiary, Recombinant Fusion Proteins, Single-Chain Antibodies |
Abstract | 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 Ca(2+)-activated gelsolin (K (d) approximately 10 nM) while GsnVHH 11 binds gelsolin irrespective of Ca(2+) (K (d) approximately 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. |
DOI | 10.1007/s00018-010-0266-1 |
Alternate Journal | Cell. Mol. Life Sci. |
PubMed ID | 20140750 |
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