F-ACTIN BINDING-SITE MASKED BY THE INTRAMOLECULAR ASSOCIATION OF VINCULIN HEAD AND TAIL DOMAINS

被引:327
作者
JOHNSON, RP [1 ]
CRAIG, SW [1 ]
机构
[1] JOHNS HOPKINS UNIV,SCH MED,DEPT BIOL CHEM,BALTIMORE,MD 21205
关键词
D O I
10.1038/373261a0
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
ALTHOUGH vinculin is present at all sites of F-actin attachment to plasma membranes(1) acid is required for linkage of myofibrils to sarcolemma(2), it is unclear how it promotes attachment of actin to membranes. Because biochemical evidence for a direct interaction of vinculin with F-actin is controversial(3-9), current models of actin-membrane linkages depict only an indirect role for vinculin, as a tether for alpha-actinin(10). We demonstrate here that an intramolecular association between the 95K head and 30K tail domains of vinculin(11) masks an F-actin binding site present in the carboxyterminal 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.
引用
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页码:261 / 264
页数:4
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