Binding and assembly of actin filaments by plasma membranes from Dictyostelium discoideum - PubMed (original) (raw)

Binding and assembly of actin filaments by plasma membranes from Dictyostelium discoideum

M A Schwartz et al. J Cell Biol. 1986 Jun.

Abstract

The binding of native, 125I-Bolton-Hunter-labeled actin to purified Dictyostelium discoideum plasma membranes was measured using a sedimentation assay. Binding was saturable only in the presence of the actin capping protein, gelsolin. In the presence of gelsolin, the amount of actin bound at saturation to three different membrane preparations was 80, 120, and 200 micrograms/mg of membrane protein. The respective concentrations of actin at half-saturation were 8, 12, and 18 micrograms/ml. The binding curves were sigmoidal, indicating positive cooperativity at low actin concentrations. This cooperativity appeared to be due to actin-actin associations during polymerization, since phalloidin converted the curve to a hyperbolic shape. In kinetic experiments, actin added as monomers bound to membranes at a rate of 0.6 microgram ml-1 min-1, while pre-polymerized actin bound at a rate of 3.0 micrograms ml-1 min-1. Even in the absence of phalloidin, actin bound to membranes at concentrations well below the normal critical concentration. This membrane-bound actin stained with rhodamine-phalloidin and was cross-linked by m-maleimidobenzoyl succinimide ester, a bifunctional cross-linker, into multimers with the same pattern observed for cross-linked F-actin. We conclude that D. discoideum plasma membranes bind actin specifically and saturably and that these membranes organize actin into filaments below the normal critical concentration for polymerization. This interaction probably occurs between multiple binding sites on the membrane and the side of the actin filament, and may be related to the clustering of membrane proteins.

PubMed Disclaimer

Cited by

Fluorescent phallotoxins as probes for filamentous actin.
Faulstich H, Zobeley S, Rinnerthaler G, Small JV. Faulstich H, et al. J Muscle Res Cell Motil. 1988 Oct;9(5):370-83. doi: 10.1007/BF01774064. J Muscle Res Cell Motil. 1988. PMID: 3063723 Review. No abstract available.
F-actin binds to the cytoplasmic surface of ponticulin, a 17-kD integral glycoprotein from Dictyostelium discoideum plasma membranes.
Wuestehube LJ, Luna EJ. Wuestehube LJ, et al. J Cell Biol. 1987 Oct;105(4):1741-51. doi: 10.1083/jcb.105.4.1741. J Cell Biol. 1987. PMID: 3312238 Free PMC article.
Binding of actin to liver cell membranes: the state of membrane-bound actin.
Tranter MP, Sugrue SP, Schwartz MA. Tranter MP, et al. J Cell Biol. 1991 Mar;112(5):891-901. doi: 10.1083/jcb.112.5.891. J Cell Biol. 1991. PMID: 1705560 Free PMC article.
Interaction of the cytoskeleton with the plasma membrane.
Niggli V, Burger MM. Niggli V, et al. J Membr Biol. 1987;100(2):97-121. doi: 10.1007/BF02209144. J Membr Biol. 1987. PMID: 3323521 Review. No abstract available.
Conformationally constrained analogues of diacylglycerol (DAG). 31. Modulation of the biological properties of diacylgycerol lactones (DAG-lactones) containing rigid-rod acyl groups separated from the core lactone by spacer units of different lengths.
Comin MJ, Czifra G, Kedei N, Telek A, Lewin NE, Kolusheva S, Velasquez JF, Kobylarz R, Jelinek R, Blumberg PM, Marquez VE. Comin MJ, et al. J Med Chem. 2009 May 28;52(10):3274-83. doi: 10.1021/jm900186m. J Med Chem. 2009. PMID: 19379015 Free PMC article.

References

1. EMBO J. 1984 Sep;3(9):2095-100 - PubMed
1. Biochim Biophys Acta. 1985;780(1):21-65 - PubMed
1. Exp Cell Res. 1977 Sep;108(2):452-6 - PubMed
1. FEBS Lett. 1984 Nov 19;177(2):184-8 - PubMed
1. Nature. 1970 Aug 15;227(5259):680-5 - PubMed

Binding and assembly of actin filaments by plasma membranes from Dictyostelium discoideum - PubMed (original) (raw)