Production and purification of a recombinant human 14 kDa β‐galactoside‐binding lectin

FEBS Letters

Hirabayashi, Jun²  Soma, Gen-Ichiro¹  Kasai, Ken-Ichi¹  Ayaki, Hitoshi¹ 

[1] Biotechnology Research Center, Teikyo University, Sagamiko, Kanagawa 199-01, Japan;Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199-01, Japan

关键词: Galactoside binding;    β-;    Lectin expression;    Cell differentiation;    (Human lung;    E. coli);    IPTG;    isopropyl-β-D-thiogalactopyranoside;    EDTA/PMSF/MEPBS;    2 mM EDTA;    1 mM phenylmethylsulfonyl fluoride;    4 mM 2-mercaptoethanol;    20 mM sodium phosphate;    pH 7.2;    0.15 M sodium chloride;    PVDF;    polyvinylidenedifluoride;    LPS;    lipopolysaccharide;   

DOI  :  10.1016/0014-5793(89)80711-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The cDNA for a 14 kDa human β-galactoside-binding lectin was inserted into a plasmid carrying a taq promoter, and the lectin protein was expressed in E. coli cells. The recombinant lectin was extracted from the cells and purified to apparent homogeneity by a single-step chromatography on an asialofetuin-agarose column. Subunit molecular mass (14 kDa), hemagglutinating activity and antigenicity were indistinguishable from those of the human placental lectin. Though the N-terminal of the placental lectin is blocked with an acetyl group, the recombinant lectin was found to have a free amino group. However, the N-terminal amino acid sequences were identical. The recombinant lectin was considered to have the same three-dimensional structure as the placental lectin.

【 授权许可】

Unknown