Purification methods of mammalian catechol-O-methyltransferases.
The protein purification strategies used for obtaining homogeneous rat and human soluble catechol-O-methyltransferase (S-COMT) polypeptides are reviewed. Expression and purification of recombinant rat and human S-COMT in Escherichia coli and for human S-COMT in baculevirus-infected insect cells made it possible to elucidate the S-COMT polypeptides in more detail. The application of these purification methods has allowed the crystallization of the rat S-COMT protein and the analysis of the kinetic properties of the enzyme in great detail. The availability of the pure S-COMT protein together with the structural data has also greatly enhanced the development of more potent COMT inhibitors.
Tilgmann
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C
Target Protein Laboratory, University of Helsinki, Finland.
Ulmanen
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I
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002394:Catechol O-Methyltransferase / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D010920:Placenta / Q000201:enzymology; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
D011994:Recombinant Proteins; D002394:Catechol O-Methyltransferase