Scarborough G A
J Biol Chem. 1975 Feb 10;250(3):1106-11.
The isolation and characterization of plasma membranes from a cell wall-less mutant of Neurospora crassa are described. The plasma membranes are stabilized against fragmentation and vesiculation by treatment of intact cells with concanavalin A just prior to lysis. After lysis, the concanavalin A-stabilized plasma membrane ghosts are isolated by low speed centrifugation techniques and the purified ghosts subsequently converted to vesicles by removal of the bulk of the concanavalin A. The yield of ghosts is about 50% whereas the yield of vesicles is about 20%. The isolated plasma membrane vesicles have a characteristically high sterol to phospholipid ratio, Mg2+-dependent ATPase activity and (Na+ plus K+)-stimulated Mg2+ATPase activity. Only traces of succinate dehydrogenase and 5'-nucleotidase are present in the plasma membrane preparations.
本文描述了从无细胞壁的粗糙脉孢菌突变体中分离和鉴定质膜的方法。在裂解前，通过用伴刀豆球蛋白A处理完整细胞，可使质膜稳定，防止其碎片化和形成囊泡。裂解后，通过低速离心技术分离出伴刀豆球蛋白A稳定的质膜空泡，然后通过去除大部分伴刀豆球蛋白A将纯化的空泡转化为囊泡。空泡的产量约为50%，而囊泡的产量约为20%。分离出的质膜囊泡具有特征性的高固醇与磷脂比率、Mg2+依赖性ATP酶活性以及（Na+加K+）刺激的Mg2+ATP酶活性。质膜制剂中仅存在痕量的琥珀酸脱氢酶和5'-核苷酸酶。
