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Inositol triphosphate receptors in sea urchin sperm

Published online by Cambridge University Press:  26 September 2008

Otilia Zapata
Affiliation:
Instituto de Biotecnología-UNAM, Cuernavaca, Mexico, University of Washington, Seattle, USA, Laboratorium voor Fysiologie, KULeuven, Belgium, and Departments of Biochemistry and Anatomy, University of Iowa, USA
James Ralston
Affiliation:
Instituto de Biotecnología-UNAM, Cuernavaca, Mexico, University of Washington, Seattle, USA, Laboratorium voor Fysiologie, KULeuven, Belgium, and Departments of Biochemistry and Anatomy, University of Iowa, USA
Carmen Beltraán
Affiliation:
Instituto de Biotecnología-UNAM, Cuernavaca, Mexico, University of Washington, Seattle, USA, Laboratorium voor Fysiologie, KULeuven, Belgium, and Departments of Biochemistry and Anatomy, University of Iowa, USA
Jan B. Parys
Affiliation:
Instituto de Biotecnología-UNAM, Cuernavaca, Mexico, University of Washington, Seattle, USA, Laboratorium voor Fysiologie, KULeuven, Belgium, and Departments of Biochemistry and Anatomy, University of Iowa, USA
Ji Long Chen
Affiliation:
Instituto de Biotecnología-UNAM, Cuernavaca, Mexico, University of Washington, Seattle, USA, Laboratorium voor Fysiologie, KULeuven, Belgium, and Departments of Biochemistry and Anatomy, University of Iowa, USA
Frank J. Longo
Affiliation:
Instituto de Biotecnología-UNAM, Cuernavaca, Mexico, University of Washington, Seattle, USA, Laboratorium voor Fysiologie, KULeuven, Belgium, and Departments of Biochemistry and Anatomy, University of Iowa, USA
Alberto Darszon*
Affiliation:
Instituto de Biotecnología-UNAM, Cuernavaca, Mexico, University of Washington, Seattle, USA, Laboratorium voor Fysiologie, KULeuven, Belgium, and Departments of Biochemistry and Anatomy, University of Iowa, USA
*
Dr Albert Darszon, Instituto de Biotecnología, postal 510-3 Cuernavaca, MO. 62250, Mexico. Tel: (52-73) 29 16 50. Fax: (52-73) 17 23 88. e-mail: [email protected].

Summary

Inositol 1,4,5-triphosphate (Ins(1,4,5)P3) is a second messenger that regulates Ca2+ channels in many important cell signalling pathways. In sea urchin sperm the outer investment of the egg triggers the acrosome reaction (AR) that involves Ins(1,4,5)P3 production and the opening of two Ca2+ channels. Here we have sought to identify a high-affinity Ins(1,4,5)P3 receptor in Strongylocentrotus purpuratus sperm. An Ins(1,4,5)P3 binding component was affinity-purified 12-fold from sperm extracts. It displayed similar characteristics to the Ins(1,4,5)P3 receptor from other sources: pH-dependent high affinity for Ins(1,4,5)3(KD=261 nM), a τ1/2 of association and dissociation of 50 and 40s, respectively, specificity (IC)50>5μM for Ins (1)P1, Ins(1,4)P2 and Ins(1,3,4,5 P4), and pharmacological sensitivity 10 and 100μ heparin/ml inhibited 75% and 100% binding respectively). An antibody against the carboxy-terminal of the type I Ins(1,4,5)P3 receptor of somatic cells recognised a Plasma membrane component in the sperm head and less intensely in the flagella. This antibody also recongnised a 240 kDa band from isolated head plasma membranes, and weakly in flagellar membrane. This IP3 receptor-like protein may mediate the sustained uptake of Ca2+ through the second Ca2+ chanel opened during the AR.

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Article
Copyright
Copyright © Cambridge University Press 1997

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