TY - JOUR
T1 - 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
AU - Wang, Jiu Yao
AU - Kishore, Uday
AU - Reid, Kenneth B.M.
PY - 1995/11/27
Y1 - 1995/11/27
N2 - 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.
AB - 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.
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U2 - 10.1016/0014-5793(95)01232-4
DO - 10.1016/0014-5793(95)01232-4
M3 - Article
C2 - 8521967
AN - SCOPUS:0028824437
VL - 376
SP - 6
EP - 10
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 1-2
ER -