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. 1998 Mar 23;140(6):1383–1393. doi: 10.1083/jcb.140.6.1383

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Actin-binding activity of actinin-4. (A) Direct association of actinin-4 with actin. In vitro translation product of actinin-4 was incubated with chicken gizzard actin-conjugated Sepharose 4B beads (lane 1) or Sepharose 4B beads alone (lane 2). After extensive washing, SDS-PAGE and autoradiography revealed that ³⁵S-labeled actinin-4 protein was retained only in actin-coupled beads (lane 1). Molecular masses (in kD) are shown on the left. (B) Cell lysate of human squamous cell carcinoma cell line (PC-10) was incubated with actin-conjugated (lanes 1 and 3) or control (lanes 2 and 4) Sepharose 4B beads. Bound proteins were analyzed by immunoblotting. Approximately 100-kD proteins of actinin-1 (lane 1) and actinin-4 (lane 3) were detected with monoclonal antibodies BM-75.2 and NCC-Lu-632, respectively. Molecular masses (in kD) are shown on the left.