Involvement of EF hand motifs in the Ca2+-dependent binding of the pleckstrin homology domain to phosphoinositides

Tada Aki Yamamoto, Hiroshi Takeuchi, Takashi Kanematsu, Victoria Allen, Hitoshi Yagisawa, Ushio Kikkawa, Yutaka Watanabe, Akihiko Nakasima, Matilda Katan, Masato Hirata

研究成果: Contribution to journalArticle査読

34 被引用数 (Scopus)


The pleckstrin homology (PH) domains of phospholipase C (PLC)-δ1 and a related catalytically inactive protein, p130, both bind inositol phosphates and inositol lipids. The binding to phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] by PLC-δ1 is proposed to be the critical interaction required for membrane localization to where the substrate resides; it is also required for the Ca2+-dependent activation of PLC-δ1 observed in the permeabilized cells. In the proximity of the PH domain, both PLC-δ1 and p130 possess the EF-hand domain, containing classical motifs implicated in calcium binding. Therefore, in the present study we examined whether the binding of the PH domain to Ptdlns(4,5)P2 is regulated by changes in free Ca2+ concentration within the physiological range. A Ca2+ dependent increase in the binding to PtdIns(4,5)P2 was observed with a full-length PLC-δ1, while the isolated PH domain did not show any Ca2+ dependence. However, the connection of the EF-hand motifs to the PH domain restored the Ca2+ dependent increase in binding, even in the absence of the C2 domain. The p130 protein showed similar properties to PLC-δ1, and the EF-hand motifs were again required for the PH domain to exhibit a Ca2+ dependent increase in the binding to PtdIns(4,5)P2. The isolated PH domains from several other proteins which have been demonstrated to bind PtdIns(4,5)P2 showed no Ca2+ dependent enhancement of binding. However, when present within a chimera also containing PLC-δ1 EF-hand motifs, the Ca2+ dependent binding was again observed. These results suggest that the binding of Ca2+ to the EF-hand motifs can modulate binding to PtdIns(4,5)P2 mediated by the PH domain.

ジャーナルEuropean Journal of Biochemistry
出版ステータス出版済み - 10 1 1999

All Science Journal Classification (ASJC) codes

  • 生化学


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