Purification and characterization of human C1-esterase inhibitor.
A new purification method for C1-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 100 000 and NH2-terminal asparagine. The molecule is fully active as judged by complex formation with C1s. Two of its three disulphide bridges can be easily reduced and S-carboxymethylated under non-denaturing conditions without loss of activity. However, at high dithioerythritol 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.
D000596:Amino Acids / Q000032:analysis; D011232:Chemical Precipitation; D002845:Chromatography; D003166:Complement Activating Enzymes / Q000378:metabolism; D003174:Complement C1 Inactivator Proteins / Q000097:blood* / Q000302:isolation & purification; D003173:Complement C1s; D004220:Disulfides; D004355:Drug Stability; D006801:Humans; D010084:Oxidation-Reduction; D011487:Protein Conformation
D000596:Amino Acids; D003174:Complement C1 Inactivator Proteins; D004220:Disulfides; D003166:Complement Activating Enzymes; D003173:Complement C1s