重组人半乳糖凝集素-1的亲和纯化与鉴定

Affinity purification and characterization of recombinant human galectin-1.

作者信息

Fouillit M, Lévi-Strauss M, Giudicelli V, Lutomski D, Bladier D, Caron M, Joubert-Caron R

机构信息

Biochimie Cellulaire des Hémopathies Lymphoïdes, UFR SMBH-Léonard de Vinci, Université Paris Nord, Bobigny, France.

出版信息

J Chromatogr B Biomed Sci Appl. 1998 Feb 27;706(1):167-71. doi: 10.1016/s0378-4347(97)00336-8.

DOI:10.1016/s0378-4347(97)00336-8

PMID:9544819

Abstract

Galectin-1, a polypeptidic factor that can have major effects on cell growth and apoptosis, was overexpressed in E. coli. This protein was purified to homogeneity by affinity chromatography on lactose coupled to divinylsulfone-activated agarose. The recombinant galectin-1 (rGAL1) was compared with the homologous protein purified from human brain tissue using two-dimensional electrophoresis on immobilized pH gradient (IPG-DALT). rGAL1 had a major isoelectric point of 5.4 (major pI of tissular galectin-1, 5.1) and its subunit molecular mass was 14500. Addition of rGAL1 to Jurkat T-lymphoblastoid cells induced cell death in a concentration-dependent manner.

摘要

半乳糖凝集素-1是一种可对细胞生长和凋亡产生重大影响的多肽因子，它在大肠杆菌中过表达。通过在与二乙烯砜活化琼脂糖偶联的乳糖上进行亲和层析，将该蛋白纯化至同质。使用固定化pH梯度双向电泳（IPG-DALT），将重组半乳糖凝集素-1（rGAL1）与从人脑组织中纯化的同源蛋白进行比较。rGAL1的主要等电点为5.4（组织半乳糖凝集素-1的主要等电点为5.1），其亚基分子量为14500。将rGAL1添加到Jurkat T淋巴母细胞中会以浓度依赖的方式诱导细胞死亡。