Actin assembly in electropermeabilized neutrophils: role of G-proteins

Biochemical and Biophysical Research Communications
G P DowneyS Grinstein

Abstract

Polymerization of microfilaments, one of the responses triggered in neutrophils by stimuli such as the chemoattractant N-formyl-methionyl-leucyl-phenylalanine (fMLP), involves the conversion of actin from the monomeric to the filamentous form. The exact sequence of events responsible for this conversion remains to be defined, but its susceptibility to inhibition by pertussis toxin provides indirect evidence that GTP-binding proteins (G-proteins) are involved. In this report, electropermeabilized cells were used to obtain more direct evidence of a role for G-proteins in actin assembly. Staining with 7-nitrobenz-2-oxa-1,3-diazole (NBD)-phallacidin and flow cytometry were used to monitor the formation of filamentous actin. GTP-gamma-S, a nonhydrolyzable analogue of GTP and aluminum fluoride, which in combination with GDP can activate G-proteins, stimulated actin assembly in electropermeabilized cells but had only marginal effects on intact cells. fMLP-induced actin polymerization in permeabilized cells was inhibited by pretreatment with GDP-beta-S, an analogue of GDP that stabilizes the inactive form of G-proteins. In contrast, stimulation by phorbol 12-myristate 13-acetate (PMA) was largely unaffected by GDP-3-S. These observatio...Continue Reading

References

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Citations

Jun 1, 1993·Journal of Cellular Physiology·G P DowneyC K Chan
Jun 8, 1993·Biochimica Et Biophysica Acta·S Orlowski, L M Mir
Jan 5, 2000·Journal of Immunological Methods·M Torres, T D Coates
Feb 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·G F HuB L Vallee
Jun 1, 1990·The Journal of Cell Biology·G P DowneyS Grinstein
Feb 1, 1992·The Journal of Cell Biology·G P DowneyS Grinstein
Sep 1, 1992·The Journal of Cell Biology·M H Symons, T J Mitchison
Dec 17, 2003·Current Opinion in Hematology·Gabriel Fenteany, Michael Glogauer
Jul 28, 1997·The Journal of Cell Biology·S H ZigmondP N Devreotes
Feb 1, 1991·Current Opinion in Cell Biology·Y L Wang
May 1, 1991·The American Journal of Physiology·S Grinstein, W Furuya
May 15, 1991·Biochemical and Biophysical Research Communications·I MelamedC M Roifman

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