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Using blot overlay techniques we have investigated the interaction of vinculin with α-actinin. We show that an α-actinin binding site is located in the 90 kDa vinculin head and confirm a vinculin binding site in the C-terminal rod of α-actinin, as recently reported by McGregor et al. [(1994) Biochem. J. 310, 225-233]. The isolated vinculin head binds much more strongly to α-actinin than intact vinculin...