Wolf M, Baggiolini M
Theodor Kocher Institut, Universität Bern, Switzerland.
Biochem J. 1990 Aug 1;269(3):723-8. doi: 10.1042/bj2690723.
Cytosol and membrane fractions from human neutrophils, monocytes, lymphocytes and platelets were separated by SDS/PAGE, blotted on to nitrocellulose and assayed for selective binding of phosphatidylserine (PS). Two PS-binding proteins with apparent molecular masses of 115 kDa and 100 kDa were identified in the cytosol of neutrophils, monocytes and lymphocytes. Corresponding bands along with other PS-binding proteins were detected in platelets in both cytosol and membrane fractions. These proteins were also found to bind protein kinase C (PKC) provided that PS was present. The 115 kDa and 100 kDa proteins (PS-p115/110) were partially purified from neutrophils and were used for the study of PS and PKC binding. The binding of PS did not require Ca2+ or Mg2+ and was inhibited by phosphatidic acid, by 1-alkyl-2-acetylphosphocholine and, to a lesser extent, by other lipids. The binding of PKC, however, was strictly PS- and Ca2(+)-dependent and seems to occur secondarily to PS binding.
通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳（SDS/PAGE）分离人中性粒细胞、单核细胞、淋巴细胞和血小板的胞质溶胶和膜组分，将其印迹到硝酸纤维素膜上，并检测磷脂酰丝氨酸（PS）的选择性结合。在中性粒细胞、单核细胞和淋巴细胞的胞质溶胶中鉴定出两种表观分子量分别为115 kDa和100 kDa的PS结合蛋白。在血小板的胞质溶胶和膜组分中检测到相应条带以及其他PS结合蛋白。还发现这些蛋白在有PS存在的情况下能结合蛋白激酶C（PKC）。从中性粒细胞中部分纯化出115 kDa和100 kDa的蛋白（PS-p115/110），并用于研究PS和PKC的结合。PS的结合不需要Ca2+或Mg2+，并受到磷脂酸、1-烷基-2-乙酰磷脂酰胆碱的抑制，其他脂质的抑制作用较小。然而，PKC的结合严格依赖于PS和Ca2+，似乎是在PS结合之后发生的。
