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SNAP-23 is not essential for constitutive exocytosis in HeLa cells
Authors:Okayama Miki  Arakawa Toshiya  Mizoguchi Itaru  Tajima Yoshifumi  Takuma Taishin
Affiliation:Department of Orthodontics, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061-0293, Japan.
Abstract:We applied the small interfering RNA (siRNA) technique and over-expression of a dominant-negative mutant to evaluate the role of SNAP-23, a non-neuronal isoform of SNAP-25, in constitutive exocytosis from HeLa cells. Although the protein level of SNAP-23 was reduced to less than 10% of the control value by siRNA directed against SNAP-23, exocytosis of SEAP (secreted alkaline phosphatase) was normal. Double knockdown of SNAP-23 and syntaxin-4 also failed to inhibit the secretion. Furthermore, over-expression of deltaC8-SNAP-23, a dominant-negative mutant of SNAP-23, did not abrogate SEAP secretion. These results suggest that SNAP-23 is not essential for constitutive exocytosis of SEAP.
Keywords:siRNA, small interfering RNA   GFP, green fluorescent protein   NSF, N-ethylmaleimide-sensitive fusion protein   SNAP, soluble NSF attachment protein   SNARE, SNAP receptor   VAMP, vesicle-associated membrane protein   SEAP, secreted alkaline phosphatase (human placental alkaline phosphatase)
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