Minnifield N, Creek K E, Navas P, Morré D J
Eur J Cell Biol. 1986 Oct;42(1):92-100.
To delineate the traffic route through the Golgi apparatus followed by newly synthesized lysosomal enzymes, we subfractionated the Golgi apparatus of rat liver by preparative free-flow electrophoresis into cisternae fractions of increasing content of trans face markers and decreasing contents of markers for the cis face. NADPase was used to mark median cisternae. Beta-Hexosaminidase, the high mannose oligosaccharide processing enzyme, alpha-mannosidase II, the two enzymes involved in the biosynthesis of the phosphomannosyl recognition marker, and the phosphomannosyl receptor itself decreased in specific activity or amount from cis to trans. Additionally, these activities were observed in a fraction consisting predominantly of cisternae, vesicles and tubules derived from trans-most Golgi apparatus elements. These results, along with preliminary pulse-labeling kinetic data for the phosphomannosyl receptor, suggest that lysosomal enzymes enter the Golgi apparatus at the cis face, are phosphorylated, and appear in trans face vesicles by a route whereby the phosphomannosyl receptor bypasses at least some median and/or trans Golgi apparatus cisternae.
为了描绘新合成的溶酶体酶在高尔基体中的运输途径，我们通过制备性自由流动电泳将大鼠肝脏的高尔基体亚分级，得到反面膜标记物含量增加而顺面膜标记物含量减少的潴泡分级。用NADP酶标记中间潴泡。β-己糖胺酶（高甘露糖寡糖加工酶）、α-甘露糖苷酶II（参与磷酸甘露糖识别标记生物合成的两种酶）以及磷酸甘露糖受体本身的比活性或含量从顺面到反面逐渐降低。此外，在一个主要由源自高尔基体最反面元件的潴泡、小泡和小管组成的分级中观察到了这些活性。这些结果，连同磷酸甘露糖受体的初步脉冲标记动力学数据，表明溶酶体酶在顺面进入高尔基体，被磷酸化，并通过磷酸甘露糖受体绕过至少一些中间和/或反高尔基体潴泡的途径出现在反面小泡中。
