A colormetric assay for catechol-O-methyltransferase.
A series of catechol diazo dyes were synthesized and tested as substrates for the enzyme catechol-O-methyltransferase (COMT) with the aim of developing a sensitive HPLC assay method using visible wavelength light detection. A method was developed which allowed for the determination of the two regioisomeric methylated products of the COMT catalyzed reaction of 4-[(3,4-dihydroxyphenyl)azo]benzenesulfonate with S-adenosylmethionine (AdoMet). Separation of the assay components was achieved by reverse phase chromatography using an isocratic mobile phase. No pre-preparation of the assay samples was required.
Bailey
Karl
K
Department of Chemistry, University of Otago, PO Box 56, Dunedin, New Zealand.
Cowling
Rebecca
R
Tan
Eng Wui
EW
Webb
Daniel
D
D001391:Azo Compounds / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D002394:Catechol O-Methyltransferase / Q000032:analysis* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003124:Colorimetry / Q000379:methods*; D004396:Coloring Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D012680:Sensitivity and Specificity; D013057:Spectrum Analysis; D013379:Substrate Specificity
D001391:Azo Compounds; D004396:Coloring Agents; D002394:Catechol O-Methyltransferase