The analysis of protein-protein interaction with special reference to PRIP-1.

• KANEMATSU Takashi

  Laboratory of Molecular Cellular Biochemistry, Faculty of Dental Science, and Station for Collaborative Research, Kyushu University

• HIRATA Masato

  Laboratory of Molecular Cellular Biochemistry, Faculty of Dental Science, and Station for Collaborative Research, Kyushu University

Bibliographic Information

Other Title

• タンパク質間相互作用解析のアプローチ
• 実験技術 タンパク質間相互作用解析のアプローチ
• ジッケン ギジュツ タンパクシツ カン ソウゴ サヨウ カイセキ ノ アプローチ

Abstract

Analysis of protein-protein interaction is one of the powerful methods for elucidating the new functions of functionally unknown proteins. Using this approach, we isolated two proteins interacting with PRIP-1, which was isolated as a new Ins(1,4,5)P₃ binding protein from brain. One was protein phosphatase 1 catalytic subunit (PP1c) and the other was GABARAP (GABA_A-receptor-associated protein). The region of PRIP-1 responsible for their interaction was the site preceding to the pleckstrin homology domain of PRIP-1 for PP1c and the EF-hand motifs of PRIP-1 for GABARAP, which were determined by β-galactosidase assay of yeast two-hybrid system. The association between PRIP-1 and PP1c was confirmed in vitro by a pull-down assay, a far-western assay, an immunoprecipitation analysis and a surface plasmon resonance analysis. The interaction of PRIP-1 with PP1c resulted in inhibition of the catalytic activity of PP1c in a PRIP-1 concentration-dependent manner. The association between PRIP-1 and GABARAP was also confirmed by a pull-down assay, and we found that PRIP-1 competitively inhibited the binding of the γ2 subunit of the GABA_A receptor to GABARAP in vitro. Our electrophysiological and behavioral analysis of PRIP-1 knockout mice revealed that PRIP-1 is essential for the function of GABA_A receptors, especially in response to the agents acting on the γ2 subunit.<br>

Journal

• Folia Pharmacologica Japonica

  Folia Pharmacologica Japonica 119 (4), 241-246, 2002

  The Japanese Pharmacological Society

Keywords

• protein-protein interaction
• PRIP-1
• PP1
• GABA_A receptor
• GABARAP

Details

• CRID: 1390001204271604096
• NII Article ID: 10008991960
• NII Book ID: AN00198335
• DOI: 10.1254/fpj.119.241
• COI: 1:STN:280:DC%2BD383kt1Kjsw%3D%3D
• ISSN: 13478397
• ISSN: 00155691
• NDL BIB ID: 6120823
• PubMed: 11979730
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I6120823
• Web Site: https://search.jamas.or.jp/link/ui/2002212501
• Text Lang: ja
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• Abstract License Flag: Disallowed