Expression of SNARE proteins in enteroendocrine cell lines and functional role of tetanus toxin‐sensitive proteins in cholecystokinin release

FEBS Letters

Takahashi, Masami²  Chayvialle, Jean-Alain¹  Regazzi, Romano³  Cuber, Jean-Claude¹  Catsicas, Stefan³  Abello, Jacques¹  Bernard, Christine¹  Bosshard, Arlette¹  Némoz-Gaillard, Eric¹ 

[1] INSERM Unité 45, Hôpital Edouard-Herriot, Pavillon Hbis, 69437 Lyon Cédex 3, France;Mitsubishi Kasei Institute of Life Sciences, Machida, Tokyo 194, Japan;Institut de Biologie Cellulaire et de Morphologie, Rue du Bugnon 9, 1005 Lausanne, Switzerland

关键词: Syntaxin;    Synaptosomal-associated protein of 25 kDa;    Vesicle-associated membrane protein;    Tetanus toxin;    Enteroendocrine cell;    Cholecystokinin exocytosis;    CCK;    cholecystokinin;    SNAP-25;    synaptosomal-associated protein of 25 kDa;    VAMP;    vesicle-associated membrane protein;    SL-O;    streptolysin-O;    Tetx;    tetanus toxin;   

DOI  :  10.1016/S0014-5793(98)00209-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In neurons, synaptic vesicle exocytosis involves the formation of a core complex particle including syntaxin-1, synaptosomal-associated protein of 25 kDa (SNAP-25) and vesicle-associated membrane protein (VAMP)-2/synaptobrevin. The expression of these proteins was investigated in a panel of cell lines, including lines of endocrine and intestinal origin, by Western blotting and/or immunocytochemistry. The three core complex proteins were detected in the enteroendocrine, cholecystokinin (CCK)-secreting, cell lines STC-1 and GLUTag, and in the endocrine non-intestinal cell lines CA-77 and HIT-T15. In contrast, SNAP-25 and syntaxin-1 were undetected in the intestinal non-endocrine cell lines IEC-6, HT-29 and Caco-2, whereas a slight expression of VAMP-2 was documented in IEC-6 and HT-29 cells. Co-immunoprecipitation experiments indicated that syntaxin-1, SNAP-25 and VAMP-2 were present in a complex similar to that identified in brain. In the STC-1 cell line, treatment of streptolysin-O-permeabilized cells with tetanus toxin (Tetx) selectively cleaved VAMP-2 and VAMP-3/cellubrevin, and simultaneously abolished Ca²⁺-induced CCK secretion (IC₅₀∼12 nM). These results show that endocrine cell lines of intestinal origin express syntaxin-1, SNAP-25 and VAMP-2, and suggest a key role for a Tetx-sensitive protein (for example VAMP-2 and/or VAMP-3) in the CCK secretion by STC-1 cells.

【 授权许可】

Unknown