DETERMINATION OF HUMAN SERUM ALCOHOL-DEHYDROGENASE USING ISOZYME-SPECIFIC FLUORESCENT SUBSTRATES

被引:13
作者
WIERZCHOWSKI, J [1 ]
HOLMQUIST, B [1 ]
VALLEE, BL [1 ]
机构
[1] HARVARD UNIV,SCH MED,CTR BIOCHEM & BIOPHYS SCI & MED,250 LONGWOOD AVE,BOSTON,MA 02115
关键词
D O I
10.1021/ac00026a017
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Both class I and class II alcohol dehydrogenase (ADH) activities are present in human serum. The contribution of each class can be measured using two class-specific, fluorogenic substrates, 4-methoxy-1-naphthaldehyde and 6-methoxy-2-naphthaldehyde. The former is highly selective for class I isozymes, especially those containing alpha or gamma-subunits, whereas class II (pi) ADH preferentially reduces the latter. Selective inhibition of class I ADH by 4-methylpyrazole further increases the specificity. Specificity accuracy, and precision of the assay for serum measurements have been determined. The activity of class I ADH in normal human serum is below the limit of detection of this method, i.e., < 1.0 nM/min. The activity of class II ADH in normal individuals is 15 +/- 5 nM/min. In some patients values as high as 2100 nM/min are observed for class I, but in all instances, the amount of class II found was higher than that of class I ADH.
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页码:181 / 186
页数:6
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