Pohlmann R, Klein U, Fromme H G, von Figura K
Hoppe Seylers Z Physiol Chem. 1981 Sep;362(9):1199-207. doi: 10.1515/bchm2.1981.362.2.1199.
Subcellular fractions of rat liver were obtained by differential centrifugation. The fractions enriched in lysosomes or microsomes were further fractionated in discontinuous sucrose density gradients or continuous iso-osmotic gradients made of modified colloidal silica. The fractions were analyzed for marker enzymes of the different subcellular organelles and for acetyl-CoA: alpha-glucosaminide N-acetyltransferase. The acetyl-CoA: alpha-glucosaminide N-acetyltransferase activity showed a bimodal distribution. About one fourth of the activity was associated with lysosomes, whereas the greater part of the activity was recovered in the microsomal fraction. Plasma membrane-enriched fractions contained only trace amounts of acetyl-CoA: alpha-glucosaminide N-acetyltransferase. The lysosomal and microsomal acetyl-CoA: alpha-glucosaminide N-acetyltransferases are membrane-bound and can be solubilized with Triton X-100. The pH dependence and sensitivity to various ions was similar for the lysosomal and microsomal enzyme.
通过差速离心获得大鼠肝脏的亚细胞组分。富含溶酶体或微粒体的组分进一步在由改良胶体二氧化硅制成的不连续蔗糖密度梯度或连续等渗梯度中进行分级分离。对这些组分进行不同亚细胞器标记酶以及乙酰辅酶A：α-氨基葡萄糖苷N-乙酰转移酶的分析。乙酰辅酶A：α-氨基葡萄糖苷N-乙酰转移酶活性呈双峰分布。约四分之一的活性与溶酶体相关，而大部分活性在微粒体组分中回收。富含质膜的组分仅含有痕量的乙酰辅酶A：α-氨基葡萄糖苷N-乙酰转移酶。溶酶体和微粒体的乙酰辅酶A：α-氨基葡萄糖苷N-乙酰转移酶是膜结合的，可用 Triton X-100 溶解。溶酶体和微粒体酶的pH依赖性和对各种离子的敏感性相似。
