Advanced search
1 file | 953.98 KB

A llama-derived gelsolin single-domain antibody blocks gelsolin-G-actin interaction

Author
Organization
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 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.
Keywords
CELL INVASION, IN-VITRO, PLASMA GELSOLIN, BINDING PROTEINS, CRYSTAL-STRUCTURE, Cancer cell, Intrabody, Cytoskeleton, Immunomodulation, Calcium, Gelsolin, Single-domain antibody, VHH, COMPLEX, APOPTOSIS, INTRABODIES, ACTIVATION, NUCLEATION

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 953.98 KB

Citation

Please use this url to cite or link to this publication:

Chicago
Van den Abbeele, Anske, Sarah De Clercq, Ariane De Ganck, Veerle De Corte, Berlinda Van Loo, Sameh Hamdy Soror, Vasundara Srinivasan, Jan Steyaert, Joël Vandekerckhove, and Jan Gettemans. 2010. “A Llama-derived Gelsolin Single-domain Antibody Blocks gelsolin-G-actin Interaction.” Cellular and Molecular Life Sciences 67 (9): 1519–1535.
APA
Van den Abbeele, A., De Clercq, S., De Ganck, A., De Corte, V., Van Loo, B., Soror, S. H., Srinivasan, V., et al. (2010). A llama-derived gelsolin single-domain antibody blocks gelsolin-G-actin interaction. CELLULAR AND MOLECULAR LIFE SCIENCES, 67(9), 1519–1535.
Vancouver
1.
Van den Abbeele A, De Clercq S, De Ganck A, De Corte V, Van Loo B, Soror SH, et al. A llama-derived gelsolin single-domain antibody blocks gelsolin-G-actin interaction. CELLULAR AND MOLECULAR LIFE SCIENCES. 2010;67(9):1519–35.
MLA
Van den Abbeele, Anske et al. “A Llama-derived Gelsolin Single-domain Antibody Blocks gelsolin-G-actin Interaction.” CELLULAR AND MOLECULAR LIFE SCIENCES 67.9 (2010): 1519–1535. Print.
@article{950959,
  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 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.},
  author       = {Van den Abbeele, Anske and De Clercq, Sarah and De Ganck, Ariane and De Corte, Veerle and Van Loo, Berlinda and Soror, Sameh Hamdy and Srinivasan, Vasundara and Steyaert, Jan and Vandekerckhove, Joël and Gettemans, Jan},
  issn         = {1420-682X},
  journal      = {CELLULAR AND MOLECULAR LIFE SCIENCES},
  keywords     = {CELL INVASION,IN-VITRO,PLASMA GELSOLIN,BINDING PROTEINS,CRYSTAL-STRUCTURE,Cancer cell,Intrabody,Cytoskeleton,Immunomodulation,Calcium,Gelsolin,Single-domain antibody,VHH,COMPLEX,APOPTOSIS,INTRABODIES,ACTIVATION,NUCLEATION},
  language     = {eng},
  number       = {9},
  pages        = {1519--1535},
  title        = {A llama-derived gelsolin single-domain antibody blocks gelsolin-G-actin interaction},
  url          = {http://dx.doi.org/10.1007/s00018-010-0266-1},
  volume       = {67},
  year         = {2010},
}

Altmetric
View in Altmetric
Web of Science
Times cited: