CAJ | 학술논문

Syntaxin 1A(Syn1A)和Munc18a蛋白在囊泡转运和分泌中起着至关重要的作用,然而它们在细胞中分选和转运的分子机制目前尚不清楚.我们用绿色荧光蛋白(EGFP)和红色荧光蛋白(TDimer2)分别标记Syn1A和Munc18a,并用荧光显微技术观察它们在BHK-21和HEK293细胞中的转运和定位.实验结果表明Syn1A主要定位在细胞质膜上,而Munc18a主要分布在胞浆中,但是与Syn1A共表达时能定位到细胞质膜上.删除胞浆部分的Syn1A蛋白不能上膜,提示其胞浆结构域在分选和定位过程中起着重要的作用.
Syntaxin 1A(Syn1A)화Munc18a단백재낭포전운화분비중기착지관중요적작용,연이타문재세포중분선화전운적분자궤제목전상불청초.아문용록색형광단백(EGFP)화홍색형광단백(TDimer2)분별표기Syn1A화Munc18a,병용형광현미기술관찰타문재BHK-21화HEK293세포중적전운화정위.실험결과표명Syn1A주요정위재세포질막상,이Munc18a주요분포재포장중,단시여Syn1A공표체시능정위도세포질막상.산제포장부분적Syn1A단백불능상막,제시기포장결구역재분선화정위과정중기착중요적작용.
Syntaxin 1A (Syn1 A) and Munc 18a play essential roles in vesicular trafficking and exocytosis. The molecular mechanism underlying the sorting of these two proteins to their physiological sites of action remains poorly understood.Here the localization of syntaxin1A (Syn1A) and Munc18a was analyzed in baby hamster kidney (BHK-21) cells and human embryonic kidney (HEK293) cells. The rat Syn1A gene was fused to the gene encoding the enhanced green fluorescent protein (EGFP). Munc18a was labeled with the red fluorescence protein (TDimer2) at its C terminal. The proteins were expressed by transient transfection in either BHK-21 or HEK293 cells. Under fluorescence microscopy, it was shown that Syn1A was shown to be transported to the plasma membrane. While Munc18a exhibited mainly cytosolic distribution when expressed alone. However, upon coexpression with Syn1A, Munc18a is translocated to the plasma membrane. In addition, a N-terminal truncated mutant Syn1A failed to localize at the plasma membrane, suggesting that the cytoplasmic domain of Syn 1A is important for its sorting and localization.