Site-specific inhibition of myosin-mediated motility in vitro by monoclonal antibodies

Paula F. Flicker, Gary Peltz, Michael Sheetz, Peter Parham, James A. Spudich

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Monoclonal antibodies directed against seven different sites on Dictyostelium myosin (Peltz, G., J. A. Spudich, and P. Parham, 1985, J. Cell Biol., 100: 1016-1023) were tested for their ability to inhibit movement of myosin in vitro, using the Nitella-based myosinmediated bead movement assay (Sheetz, M. P., R. Chasan, and J. A. Spudich, 1984, J. Cell Biol., 99: 1867-1871). To complement this functional assay, we located the binding sites of these antibodies by electron microscopy, using the rotary shadowing technique. One antibody bound to the 18,000-dalton light chain and inhibited movement completely. All of the remaining antibodies bound to various positions along the rod portion of the myosin molecule, which is ≈1,800 A long. Antibodies that bound to the rod about 470, 680, and 1400 A from the head-tail junction did not alter myosin movement. One antibody appeared to bind very close to the head-tail junction and to inhibit movement 50%. Surprisingly, three antibodies that bound about 1,200 A from the head-tail junction inhibited movement completely. This inhibition did not depend on using intact IgG, since Fab’ fragments had the same effect.

Original languageEnglish (US)
Pages (from-to)1024-1030
Number of pages7
JournalJournal of Cell Biology
Volume100
Issue number4
DOIs
StatePublished - Apr 1 1985
Externally publishedYes

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Myosins
Monoclonal Antibodies
Antibodies
Head
Nitella
Antibody Binding Sites
Myosin Subfragments
Gastrin-Secreting Cells
Immunoglobulin Fab Fragments
Dictyostelium
Electron Microscopy
Immunoglobulin G
In Vitro Techniques
Light

ASJC Scopus subject areas

  • Cell Biology

Cite this

Site-specific inhibition of myosin-mediated motility in vitro by monoclonal antibodies. / Flicker, Paula F.; Peltz, Gary; Sheetz, Michael; Parham, Peter; Spudich, James A.

In: Journal of Cell Biology, Vol. 100, No. 4, 01.04.1985, p. 1024-1030.

Research output: Contribution to journalArticle

Flicker, Paula F. ; Peltz, Gary ; Sheetz, Michael ; Parham, Peter ; Spudich, James A. / Site-specific inhibition of myosin-mediated motility in vitro by monoclonal antibodies. In: Journal of Cell Biology. 1985 ; Vol. 100, No. 4. pp. 1024-1030.
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