An interaction between zyxin and alpha-actinin.
Zyxin is an 82-kD protein first identified as a component of adhesion plaques and the termini of stress fibers near where they associate with the cytoplasmic face of the adhesive membrane. We report here that zyxin interacts with the actin cross-linking protein alpha-actinin. Zyxin cosediments with filamentous actin in an alpha-actinin-dependent manner and an association between zyxin and alpha-actinin is observed in solution by analytical gel filtration. The specificity of the interaction between zyxin and alpha-actinin was demonstrated by blot overlay experiments in which 125I-zyxin recognizes most prominently alpha-actinin among a complex mixture of proteins extracted from avian smooth muscle. By these blot overlay binding studies, we determined that zyxin interacts with the NH2-terminal 27-kD domain of alpha-actinin, a region that also contains the actin binding site. Solid phase binding assays were performed to evaluate further the specificity of the binding and to determine the affinity of the zyxin-alpha-actinin interaction. By these approaches we have demonstrated a specific, saturable, moderate-affinity interaction between zyxin and alpha-actinin. Furthermore, double-label immunofluorescence reveals that zyxin and alpha-actinin exhibit extensive overlap in their subcellular distributions in both chicken embryo fibroblasts and pigmented retinal epithelial cells. The significant colocalization of the two proteins is consistent with the possibility that the interaction between zyxin and alpha-actinin has a biologically relevant role in coordinating membrane-cytoskeletal interactions.
Crawford
A W
AW
Department of Biology, University of Utah, Salt Lake City 84112.
Michelsen
J W
JW
Beckerle
M C
MC
D000185:Actinin / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000378:metabolism*; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002642:Chick Embryo; D002850:Chromatography, Gel; D005455:Fluorescent Antibody Technique; D007365:Intercellular Junctions / Q000737:chemistry*; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D010857:Pigment Epithelium of Eye
D015815:Cell Adhesion Molecules; D008565:Membrane Proteins; D000185:Actinin