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Vol. 12, Issue 5, 1227-1237, May 2001
v
3-Vitronectin Binding Is a
Multistage Process in which Increases in Bond Strength Are Dependent on
Y747 and Y759 in the Cytoplasmic Domain of 3
and
Integrin receptors serve as mechanical links between the
cell and its structural environment. Using Department of Microbiology, University of Pennsylvania,
Philadelphia, Pennsylvania 19104; and Department of Cell and
Developmental Biology at the State University of New York, Upstate
Medical University, Syracuse, New York 12310
v
3
integrin expressed in K562 cells as a model system, the process
by which the mechanical connection between v3 and
vitronectin develops was analyzed by measuring the resistance of these
bonds to mechanical separation. Three distinct stages of activation, as
defined by increases in the v3-vitronectin
binding strength, were defined by mutational, biochemical, and biomechanical analyses. Activation to the low binding
strength stage 1 occurs through interaction with the vitronectin ligand
and leads to the phosphorylation of Y747 in the 3 subunit. Stage 2 is characterized by a 4-fold increase in binding strength and is
dependent on stage1 and the phosphorylation of Y747. Stage 3 is
characterized by a further 2.5-fold increase in binding strength and is
dependent on stage 2 events and the availability of Y759 for
interaction with cellular proteins. The Y747F mutant blocked the
transition from stage 1 to stage 2, and the Y759F blocked the
transition from stage 2 to stage 3. The data suggest a model for
tension-induced activation of v3 integrin.
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