論文

査読有り
2012年12月

Constitutive Turnover of Phosphorylation at Thr-412 of Human p57/Coronin-1 Regulates the Interaction with Actin

JOURNAL OF BIOLOGICAL CHEMISTRY
  • Teruaki Oku
  • ,
  • Mai Nakano
  • ,
  • Yutaka Kaneko
  • ,
  • Yusuke Ando
  • ,
  • Hiroki Kenmotsu
  • ,
  • Saotomo Itoh
  • ,
  • Makoto Tsuiji
  • ,
  • Yoshiyuki Seyama
  • ,
  • Satoshi Toyoshima
  • ,
  • Tsutomu Tsuji

287
51
開始ページ
42910
終了ページ
42920
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1074/jbc.M112.349829
出版者・発行元
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

The actin-binding protein p57/coronin-1, a member of the coronin protein family, is selectively expressed in hematopoietic cells and plays crucial roles in the immune response through reorganization of the actin cytoskeleton. We previously reported that p57/coronin-1 is phosphorylated by protein kinase C, and the phosphorylation down-regulates the association of this protein with actin. In this study we analyzed the phosphorylation sites of p57/coronin-1 derived from HL60 human leukemic cells by MALDI-TOF-MS, two-dimensional gel electrophoresis, and Phos-tag (R) acrylamide gel electrophoresis in combination with site-directed mutagenesis and identified Ser-2 and Thr-412 as major phosphorylation sites. A major part of p57/coronin-1 was found as an unphosphorylated form in HL60 cells, but phosphorylation at Thr-412 of p57/coronin-1 was detected after the cells were treated with calyculin A, a Ser/Thr phosphatase inhibitor, suggesting that p57/coronin-1 undergoes constitutive turnover of phosphorylation/dephosphorylation at Thr-412. A diphosphorylated form of p57/coronin-1 was detected after the cells were treated with phorbol 12-myristate 13-acetate plus calyculin A. We then assessed the effects of phosphorylation at Thr-412 on the association of p57/coronin-1 with actin. A co-immunoprecipitation experiment with anti-p57/coronin-1 antibodies and HL60 cell lysates revealed that beta-actin was co-precipitated with the unphosphorylated form but not with the phosphorylated form at Thr-412 of p57/coronin-1. Furthermore, the phosphorylation mimic (T412D) of p57/coronin-1 expressed in HEK293T cells exhibited lower affinity for actin than the wild-type or the unphosphorylation mimic (T412A) did. These results indicate that the constitutive turnover of phosphorylation at Thr-412 of p57/coronin-1 regulates its interaction with actin.

Web of Science ® 被引用回数 : 6

リンク情報
DOI
https://doi.org/10.1074/jbc.M112.349829
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/23100250
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000312501600049&DestApp=WOS_CPL
ID情報
  • DOI : 10.1074/jbc.M112.349829
  • ISSN : 0021-9258
  • PubMed ID : 23100250
  • Web of Science ID : WOS:000312501600049

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