从小鼠肺中快速纯化两种脂皮质素样蛋白。

Rapid purification of two lipocortin-like proteins from mice lung.

作者信息

Errasfa M, Russo-Marie F

机构信息

Unité associée INSERM n. 285/Institut Pasteur, Paris, France.

出版信息

Biochem Biophys Res Commun. 1988 Jun 30;153(3):1271-5. doi: 10.1016/s0006-291x(88)81365-2.

DOI:10.1016/s0006-291x(88)81365-2

PMID:2968796

Abstract

We have purified two proteins (40 kDa and 36 kDa) from mice lung by the method of calcium-precipitation/EGTA solubilization and then a separation on a high anion exchanger column (Mono Q HR 5/5. Pharmacia) with a gradient of NaCl. The two proteins were strong inhibitors of phospholipase A2 as assessed in vitro with porcine pancreatic phospholipase A2 and [3H]-oleic acid labeled E. Coli membranes as substrate. The 40 kDa protein had a pI of 5.8 and was found to be immunologically related to human recombinant lipocortin I. The 36 kDa protein had a pI of 4.7 and cross-reacted with a polyclonal antibody raised against a 32 kDa human lipocortin-like protein described in human blood mononuclear cells. We report here a rapid purification of two distinct lipocortin-like proteins from mice lung.

摘要

我们通过钙沉淀/乙二醇双乙胺四乙酸（EGTA）增溶法从小鼠肺中纯化了两种蛋白质（40 kDa和36 kDa），然后在高阴离子交换柱（Mono Q HR 5/5，Pharmacia）上用氯化钠梯度进行分离。用猪胰磷脂酶A2和[3H] -油酸标记的大肠杆菌膜作为底物进行体外评估时，这两种蛋白质是磷脂酶A2的强效抑制剂。40 kDa的蛋白质的等电点为5.8，并且发现其与人类重组脂皮质素I存在免疫相关性。36 kDa的蛋白质的等电点为4.7，并且与针对人血单核细胞中描述的一种32 kDa人脂皮质素样蛋白产生的多克隆抗体发生交叉反应。我们在此报告从小鼠肺中快速纯化出两种不同的脂皮质素样蛋白质。