Two Regions Responsible for the Actin Binding of p57, a Mammalian Coronin Family Actin-Binding Protein

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概要

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• The aetin-binding protein p57, a member of the coronin protein family, is expressed in a variety of immune cells. It has five WD repeats and a coiled-coil motif containing a leucine zipper, both of which are known to mediate protein-protein interactions. In order to identify the precise actin-binding regions in p57, and to assess the contribution of these structural motifs, we prepared various truncated p57 as fusion proteins with glutathione S-transferase (GST) and examined their actin-binding activity. A co-sedimentation assay demonstrated that p57^<1〜371> (C-terminal truncated p57) had the ability to bind F-actin, but p57^<372-461> (a fragment containing the coiled-coil motif) did not. A segment consisting of the N-terminal 34 amino acids of p57 (p57^<1-34>) was found to bind to F-actin in the co-sedimentation assay. Furthermore, fluorescence microscopic observation showed that p57^<1-34> was co-localized with F-actin in COS-1 cells after the transfection with the p57^<1-34> construct. Deletion of ^<10>KFRHVF^<15>, a sequence conserved among coronin-related proteins, from p57^<1-34> abolished its actin-binding activity, suggesting that this sequence with basic and hydrophobic amino acids is crucial for p57 to bind to F-actin. However, the N-terminal deletion mutant p57^<63-461> retained the binding ability to F-actin. This result suggests the presence of a second actin-binding region. Further deletion analysis revealed that p57^<111-204>, which includes the second and third WD repeats, also exhibited weak actin-binding activity in the co-sedimentation assay. Taken together, these data strongly suggest that at least two regions within Met-1 to Asp-34 and Ile-111 to Glu-204 of p57 are responsible for its binding to the actin cytoskeleton.

• 公益社団法人日本薬学会の論文
• 2003-04-01

著者

• 鈴木 和美

  Showa College of Pharmaceutical Sciences

• Suzuki Kazutoshi

  放射線医学総合研究所分子イメージング研究センター

• Suzuki K

  Pharmaceutical Frontier Research Laboratories Japan Tobacco Inc.

• Suzuki Kenji

  Suntory Institute For Biomedical Research Wakayamadai

• Suzuki K

  Noda Research Laboratories Drug Delivery System Institute Ltd.:(present Address)pharmaceutics Resear

• Kato Shingo

  Research Center Hospital For Charged Particle Therapy National Institute Of Radiological Sciences

• TOYOSHIMA Satoshi

  Pharmaceuticals and Medical Devices Evaluation Center National Institute of Health Sciences

• Oku Teruaki

  Department Of Microbiology Hoshi University School Of Pharmacy And Pharmaceutical Sciences

• Oku Teruaki

  Department Of Microbiology School Of Pharmacy Hoshi University

• OKU Teruaki

  Hoshi University School of Pharmacy and Pharmaceutical Sciences

• ITOH Saotomo

  Hoshi University School of Pharmacy and Pharmaceutical Sciences

• OKUNO Masamitsu

  Hoshi University School of Pharmacy and Pharmaceutical Sciences

• SUZUKI Akio

  Hoshi University School of Pharmacy and Pharmaceutical Sciences

• SUZUKI Kensuke

  Pharmaceutical Frontier Research Laboratories, Japan Tobacco, Inc.

• NAKAJIN Shizuo

  Hoshi University School of Pharmacy and Pharmaceutical Sciences

• NAUSEEF William

  The Inflammation Program and Department of Medicine, University of Iowa and Veterans Affairs Medical

• Suzuki Kazutoshi

  Department Of Medical Imaging National Institute Of Radiological Sciences

• Suzuki Kazutoshi

  Clinical Neuroimaging Section Department Of Molecular Neuroimaging Molecular Imaging Center National

• Suzuki Kensuke

  Pharmaceutical Frontier Research Laboratories Japan Tobacco Inc.

• Toyoshima S

  Department Of Biochemistry Faculty Of Pharmaceutical Sciences Hoshi University

• Toyoshima S

  Pharmaceuticals And Medical Devices Agency

• Toyoshima Satoshi

  Pharmaceutical And Medical Devices Evaluation Center National Institute Of Health Science

• Suzuki A

  Tohoku Pharmaceutical Univ. Sendai Jpn

• Suzuki K

  Molecular Imaging Center National Institute Of Radiological Sciences

• Oku T

  Amagasaki Chemical Industries Co.

• Nakajin S

  Hoshi University School Of Pharmacy And Pharmaceutical Sciences

• Nakajin Shizuo

  Hoshi College Of Pharmacy

• Nauseef William

  The Inflammation Program And Department Of Medicine University Of Iowa And Veterans Affairs Medical

• Itoh Saotomo

  Department Of Microbiology Hoshi University School Of Pharmacy And Pharmaceutical Sciences

• Sakane T

  St. Marianna Univ. School Of Medicine Kanagawa Jpn

• Okano Masamitsu

  Hoshi University School of Pharmacy and Pharmaceutical Sciences

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