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Synaptotagmin(Syt)constitutes a family of membrane-trafficking proteins,so far nearly 20 Syts have beendiscovered.Extensive work showed that synatotagmins were a potential Ca~(2+) sensor for regulated exocytosis.Thisstudy was to investigate the expression and location of synaptotagmin Ⅱ(Syt2)in RBL-2H3(RBL)and its role inregulating exocytosis of RBL.The expression of Syt2 in RBL was confirmed by Western blot.The recombinantexpression vector pEGFP-N1-Syt2 was constructed and transfected into RBL by electroporation,the stabletransfectant RBL-Syt2-S expressing fusion protein Syt2-EGFP were obtained and Syt2 was highly concentrated atplasma membrane with little detected in cytoplasm.To analyze the role of Syt2 during exocytosis of RBL,therelease of cathepsin D was assayed by immunoblotting.Compared with control,the release of cathepsin D byRBL-Syt2-S was markedly decreased.The results indicated that Syt2 played a negative regulation in exocytosis oflysosomes in RBL.Cellular & Molecular Immunology.2005;2(3):205-209.
Synaptotagmin (Syt) constitutes a family of membrane-trafficking proteins, so far nearly 20 Syts have beendiscovered. Extensive work showed that synatotagmins were a potential Ca ~ (2+) sensor for regulated exocytosis. This study was to investigate the expression and location of synaptotagmin The expression of Syt2 in RBL was confirmed by Western blot. The recombinant plasmid pEGFP-N1-Syt2 was constructed and transfected into RBL by electroporation, the stable transfectant RBL-Syt2-S expressing fusion protein Syt2-EGFP were obtained and Syt2 was highly concentrated atplasma membrane with little detected in cytoplasm. To analyze the role of Syt2 during exocytosis of RBL, therelease of cathepsin D was assayed by immunoblotting. Compared with control, the release of cathepsin D byRBL-Syt2-S was markedly decreased.The results indicated that Syt2 played a negative regulation in exocytosis of lysosomes in RBL. Cellular & Molecular I mmunology. 2005; 2 (3): 205-209.