A recombinant polypeptide, composed of the α‐helical neck region and the carbohydrate recogniton domain of conglutinin, self‐associates to give a functionally intact homotrimer

FEBS Letters

Kishore, Uday¹  Reid, Kenneth B.M.¹  Wang, Jiu-Yao¹ 

[1] MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK

关键词: Collectin;    Recombinant bovine conglutinin;    Carbohydrate recognition domain;    Lipopolysaccharide;    C3 component of complement;    C-type lectin;   

DOI  :  10.1016/0014-5793(95)01232-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A recombinant polypeptide composed of the α-helical neck region and carbohydrate recognition domain (CRD) of bovine conglutinin was expressed in Escherichia coli. The recombinant protein formed inclusion bodies but could be solubilised using a denaturation-renaturation cycle based on urea and then purified by affinity chromatography on a TSK-N-acetylglucosamine column. The purified product behaved as a homotrimer in nondissociating conditions, with three CRDs held together by the α-helical neck regions. The trimer, although lacking the N-terminal and collagen regions of the native conglutinin, showed the same binding carbohydrate specificities as the native molecule, for the complement fragment C3b and for lipopolysaccharides derived from Gram-negative bacteria.

【 授权许可】

Unknown