Purification of rat liver soluble catechol-O-methyltransferase by high performance liquid chromatography

✍ Scribed by Tapio Korkolainen; Erkki Nissinen

• Publisher: John Wiley and Sons
• Year: 1989
• Tongue: English
• Weight: 324 KB
• Volume: 3
• Category: Article
• ISSN: 0269-3879
• DOI: 10.1002/bmc.1130030308

✦ Synopsis

The soluble form of catechol-0-methyltransferase (EC 2.1.1.6.) from rat liver was purified to homogeneity by high-performance anion-exchange chromatography and high-performance gel-filtration chromatography. The specific activity of the final pool was 270 U/mg protein. The purification was 1180-fold and recovery of the enzyme activity was 15%. During this rapid and gentle purification there were no problems with loss of activity, and the estimated half life of the final purified enzyme pool was 5.5 days at +4 "C. The only additive used was phenylmethylsulfonylfluoride in the homogenizing buffer.