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Purification and characterization of human ci-esterase inhibitor

1982; Elsevier BV; Volume: 705; Issue: 2 Linguagem: Inglês

10.1016/0167-4838(82)90188-1

1878-1454

Torbjörn Nilsson, Björn Wiman,

Enzyme function and inhibition

A new purification method for Cl̄-esterase inhibitor is described, which is essentially a three-step procedure: precipitation with poly(ethylene glycol), chromatography on DEAE-cellulose and hydrophobic interaction chromatography on hexyl-Sepharose. The final product is a single-chain glycoprotein with a molecular weight of about 100000 and NH2-terminal asparagine. The molecule is fully active as judged by complex formation with Cl̄s. Two of its three disulphide bridges can be easily reduced and S-carboxymethylated under non-denaturing conditions without loss of activity. However, at high dithioerythritoi concentration the third disulphide bridge is also cleaved and accompanied by loss of the activity, indicating that this disulphide bridge is involved in maintaining the conformation around the reactive site in the inhibitor.