F-actin binding site masked by the intramolecular association of vinculin head and tail domains

Robert P. Johnson, Susan W. Craig

Research output: Contribution to journalArticle

Abstract

ALTHOUGH vinculin is present at all sites of F-actin attachment to plasma membranes1 and is required for linkage of myofibrils to sarcolemma2, it is unclear how it promotes attachment of actin to membranes. Because biochemical evidence for a direct interaction of vinculin with F-actin is controversial3-9, current models of actin-membrane linkages depict only an indirect role for vinculin, as a tether for α-actinin10. We demonstrate here that an intramolecular association between the 95K head and 30K tail domains of vinculin11 masks an F-actin binding site present in the carboxy-terminal tail domain. Cosedimentation and crosslinking assays, and direct visualization by transmission electron microscopy, reveal an interaction between F-actin and a bacterially expressed fusion protein containing amino acids 811-1066 of vinculin, and between F-actin and a proteolytic fragment of vinculin containing amino acids 858-1066. Vinculin itself neither cosediments with nor crosslinks F-actin. The amino-terminal 95K head fragment of vinculin, but not intact vinculin, inhibits both cosedimentation and crosslinking. We propose that assembly of vinculin into an adherens junction involves disruption of the head-tail interaction, revealing a site that mediates microfilament attachment.

Original languageEnglish (US)
Pages (from-to)261-264
Number of pages4
JournalNature
Volume373
Issue number6511
StatePublished - 1995

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Vinculin
Actins
Binding Sites
Head
Adherens Junctions
Amino Acids
Membranes
Myofibrils
Masks
Transmission Electron Microscopy
Actin Cytoskeleton

ASJC Scopus subject areas

  • General

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F-actin binding site masked by the intramolecular association of vinculin head and tail domains. / Johnson, Robert P.; Craig, Susan W.

In: Nature, Vol. 373, No. 6511, 1995, p. 261-264.

Research output: Contribution to journalArticle

Johnson, RP & Craig, SW 1995, 'F-actin binding site masked by the intramolecular association of vinculin head and tail domains', Nature, vol. 373, no. 6511, pp. 261-264.
Johnson, Robert P. ; Craig, Susan W. / F-actin binding site masked by the intramolecular association of vinculin head and tail domains. In: Nature. 1995 ; Vol. 373, No. 6511. pp. 261-264.
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