Evidence for the activation of PAR‐2 by the sperm protease, acrosin: expression of the receptor on oocytes

FEBS Letters

Tomlinson, Jim²  Pike, Robert N.¹  Smith, Rosealee¹  Jenkins, Alison¹  Mackie, Eleanor J.⁵  Deshpande, Usha²  Jones, Roy³  Ramakrishnan, Vanitha²  Thompson, Philip⁶  Lourbakos, Afrodite¹  Johnson, David A.⁴ 

[1] Department of Biochemistry and Molecular Biology, Monash University, Clayton, Vic. 3800, Australia;COR Therapeutics Inc., South San Francisco, CA 94080, USA;Department of Development and Signalling, The Babraham Institute, Cambridge CB2 4AT, UK;Department of Biochemistry and Molecular Biology, J.H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN 37614, USA;School of Veterinary Science, University of Melbourne, Parkville, Vic. 3010, Australia;Department of Medicine, Monash University, Boxhill, Vic. 3128, Australia

关键词: Protease-activated receptor-2;    Acrosin;    Fluorescent quenched substrate;    Intracellular calcium;    Oocyte;    Immunocytochemistry;    PAR-2;    proteinase-activated receptor-2;    Abz;    aminobenzoic acid;    Dnp;    dinitrophenol;    CHO cells;    Chinese hamster ovary cells;    BSA;    bovine serum albumin;    APC;    activated protein C;    [Ca2+]i;    intracellular calcium concentration;    Fura-2/AM;    Fura-2-acetoxymethyl ester;    FQS;    fluorescent quenched substrate;    AEBSF;    4-(2-aminoethyl)-benzenesulfonyl fluoride;   

DOI  :  10.1016/S0014-5793(00)02146-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Proteinase-activated receptor-2 (PAR-2) is a member of a family of G-protein-coupled, seven-transmembrane domain receptors that are activated by proteolytic cleavage. The receptor is expressed in a number of different tissues and potential physiological activators identified thus far include trypsin and mast cell tryptase. Acrosin, a trypsin-like serine proteinase found in spermatozoa of all mammals, was found to cleave a model peptide fluorescent quenched substrate representing the cleavage site of PAR-2. This substrate was cleaved with kinetics similar to those of the known PAR-2 activators, trypsin and mast cell tryptase. Acrosin was also shown to induce significant intracellular calcium responses in Chinese hamster ovary cells stably expressing intact human PAR-2, most probably due to activation of the receptor. Immunohistochemical studies using PAR-2 specific antibodies indicated that the receptor is expressed by mouse oocytes, which suggests that acrosin may play additional role(s) in the fertilization process via the activation of PAR-2 on oocytes.

【 授权许可】

Unknown