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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
Author(s) -
Wang Jiu-Yao,
Kishore Uday,
Reid Kenneth B.M.
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01232-4
Subject(s) - recombinant dna , trimer , biochemistry , denaturation (fissile materials) , escherichia coli , affinity chromatography , chemistry , inclusion bodies , carbohydrate , urea , biology , dimer , enzyme , organic chemistry , nuclear chemistry , gene
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.