Covalent and non‐covalent interaction of chymotrypsin with α2‐macroglobulin

FEBS Letters

Delain, Etienne¹  Favaudon, Vincent²  Tourbez, Martine²  Pochon, François² 

[1] Laboratoire de Microscopie Cellulaire et Moléculaire, Institut Gustave Roussy, 94805 Villejuif, France;Unité 219 INSERM, Institut Curie-Biologie, Centre Universitaire, 91405 Orsay France

关键词: Chymotrypsin;    α2-Macroglobulin;    Covalent interaction;    Non-covalent interaction;    Electrophoresis;    α2M;    α2-macroglobuIin;    FITC;    fluorescein isothiocyanate;    I-AEDns;    N-(iodoacetylaminoethyl)-5-naphthylamine-1-sulfonate;    Hepes;    N-(2-hydroxyethyl)piperazine-N′-2-ethanesulfonate;   

DOI  :  10.1016/0014-5793(87)81251-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The pattern of covalent crosslinking between human α₂-macroglobulin (α₂M) and chymotrypsin has been investigated by chromatography and polyacrylamide gel electrophoresis in denaturing medium. Reaction with a single mol of chymotrypsin per mol α₂M results in the formation of a 95% covalent 1:1 chymotrypsin-α₂M complex and in the proteolytic cleavage of both 180 kDa monomers in one α₂M subunit. Proteolytic cleavage in the other α₂M subunit requires the presence of a second mol of chymotrypsin; part (20%) of the protease in the 2:1 chymotrypsin-α₂M complex thus formed appears to be non-covalently bound to the α₂M chains. Covalent binding is abolished when the reaction of α₂M with the protease is carried out in the presence of hydroxylamine. A single mol of the protease is then able to cleave all four 180 kDa monomers in α₂M.

【 授权许可】

Unknown