Mammalian ESCRT-III-Related Protein IST1 Has a Distinctive Met-Pro Repeat Sequence That Is Essential for Interaction with ALG-2 in the Presence of Ca²⁺

• OKUMURA Mayumi

  Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University

• TAKAHASHI Takeshi

  Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University

• SHIBATA Hideki

  Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University

• MAKI Masatoshi

  Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University

Bibliographic Information

Other Title

• Mammalian ESCRT-Ⅲ-Related Protein IST1 Has a Distinctive Met-Pro Repeat Sequence That Is Essential for Interaction with ALG-2 in the Presence of Ca²⁺

Abstract

ALG-2 is an EF-hand-type Ca²⁺-binding protein that interacts with a variety of intracellular proteins that possess Pro-rich regions (PRRs) in mammalian cells. IST1 is an endosomal sorting complex required for transport (ESCRT)-III-related charged multivesicular body protein (CHMP)-like protein, but unlike other ESCRT-III proteins, mammalian IST1 has a PRR and a distinctive sequence of Met-Pro repeats. We found that ALG-2 binds to IST1 by Far-Western analysis using biotinylated ALG-2 as probe, and that the Met-Pro repeat sequence is essential for interaction. The results of pulldown assays using Strep-tagged ALG-2 and lysates of cells expressing GFP-fused IST1 proteins indicated that the binding of ALG-2 to IST1 is Ca²⁺-dependent, and that it is enhanced by co-expression with CHMP1 proteins. Moreover, pulldown assays using various mutants of GST-ALG-2 revealed that the ability of IST1 to bind to mutants is different from those of known ALG-2-interacting proteins, suggesting that IST1 binds to ALG-2 by a different mode of recognition.

Journal

• Bioscience, Biotechnology, and Biochemistry

  Bioscience, Biotechnology, and Biochemistry 77 (5), 1049-1054, 2013

  Japan Society for Bioscience, Biotechnology, and Agrochemistry

Keywords

• Ca²⁺-binding protein
• repeats of two amino acids
• protein-protein interaction
• Pro-rich region
• pulldown assay

Details

• CRID: 1390001206477974016
• NII Article ID: 10031177595
• NII Book ID: AA10824164
• DOI: 10.1271/bbb.130022
• COI: 1:STN:280:DC%2BC3snht1Chsg%3D%3D
• ISSN: 13476947
• ISSN: 09168451
• NDL BIB ID: 024526190
• PubMed: 23649269
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I024526190
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.130022
• Web Site: https://search.jamas.or.jp/link/ui/2013339648
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed