Fishman J B, Fine R E
Cell. 1987 Jan 16;48(1):157-64. doi: 10.1016/0092-8674(87)90366-7.
We used a cholinesterase-mediated density shift protocol to investigate the movement of internalized transferrin (Tf) through endo- and exocytic coated vesicles (CVs) in the perfused rat liver. Upon internalization, exogenous 125I-Tf was found in endocytic CVs but not in cholinesterase-containing (i.e., exocytic) CVs (0-40 min). Between 1 and 2 hr, 125I-Tf began to appear in exocytic CVs. The origin of the exocytic CV was further investigated. After perfusion of the liver with asialotransferrin, the exocytic CVs were shown to contain resialylated Tf, indicating that the trans Golgi was the origin of this class of CVs. The resialylated Tf accumulated in the extracellular medium with kinetics very similar to the time course for appearance of Tf in cholinesterase-containing, exocytic CVs, suggesting that these CVs are directly involved in the transfer of material from the trans Golgi to the cell surface.
我们采用胆碱酯酶介导的密度转移实验方案,研究内化转铁蛋白(Tf)在灌注大鼠肝脏中通过内吞和外排被膜小泡(CVs)的转运情况。内化后,外源性125I-Tf出现在内吞CVs中,但在含胆碱酯酶的(即外排的)CVs中未出现(0 - 40分钟)。1至2小时之间,125I-Tf开始出现在外排CVs中。我们进一步研究了外排CVs的来源。用去唾液酸转铁蛋白灌注肝脏后,发现外排CVs含有重新唾液酸化的Tf,这表明反式高尔基体是这类CVs的来源。重新唾液酸化的Tf在细胞外介质中积累,其动力学与Tf出现在含胆碱酯酶的外排CVs中的时间进程非常相似,这表明这些CVs直接参与了物质从反式高尔基体向细胞表面的转运。