Slusarewicz P, Nilsson T, Hui N, Watson R, Warren G
Cell Biology Laboratory, Imperial Cancer Research Fund, London, United Kingdom.
J Cell Biol. 1994 Feb;124(4):405-13. doi: 10.1083/jcb.124.4.405.
Rat liver Golgi stacks were extracted with Triton X-100 at neutral pH. After centrifugation the low speed pellet contained two medial-Golgi enzymes, N-acetylglucosaminyltransferase I and mannosidase II, but no enzymes or markers from other parts of the Golgi apparatus. Both were present in the same structures which appeared, by electron microscopy, to be small remnants of cisternal membranes. The enzymes could be removed by treatment with low salt, leaving behind a salt pellet, which we term the matrix. Removal of salt caused specific re-binding of both enzymes to the matrix, with an apparent dissociation constant of 3 nM for mannosidase II. Re-binding was abolished by pretreatment of intact Golgi stacks with proteinase K, suggesting that the matrix was present between the cisternae.
在中性pH条件下，用Triton X-100提取大鼠肝脏高尔基体堆叠。离心后，低速沉淀中含有两种中间高尔基体酶，N-乙酰葡糖胺基转移酶I和甘露糖苷酶II，但没有来自高尔基体其他部分的酶或标志物。两者都存在于相同的结构中，通过电子显微镜观察，这些结构似乎是扁平囊膜的小残余物。用低盐处理可去除这些酶，留下一个盐沉淀，我们将其称为基质。去除盐会导致两种酶特异性地重新结合到基质上，甘露糖苷酶II的表观解离常数为3 nM。用蛋白酶K预处理完整的高尔基体堆叠可消除重新结合，这表明基质存在于扁平囊之间。
