SIMULTANEOUS DETERMINATION OF ENZYME-ACTIVITY AND ENZYME QUANTITY IN SINGLE HUMAN ERYTHROCYTES

被引:13
|
作者
TAN, WH [1 ]
YEUNG, ES [1 ]
机构
[1] IOWA STATE UNIV SCI & TECHNOL,DEPT CHEM,AMES,IA 50011
关键词
D O I
10.1006/abio.1995.1193
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An integrated laser-based fluorescence enzyme assay and particle-counting immunoassay system with zeptomole sensitivity has been developed for simultaneous determinations of activity and concentration of enzymes in single erythrocytes. The product NADPH of the reaction between glucose-6-phosphate and NADP(+) catalyzed by glucose-6-phosphate dehydrogenase (G6PDH) is monitored. Simultaneously, the agglutination of antibody-coated particles in the presence of G6PDH produces large particles which are counted by light scattering. The correlation between the two parameters indicates that on average only about 35% of the enzyme in individual cells is active. There is more than a 10-fold cell-to-cell variation in the ratios of the two parameters. This indicates that there are multiple mechanisms for the degradation of intracellular enzymes as a function of cell age. (C) 1995 Academic Press, Inc.
引用
收藏
页码:74 / 79
页数:6
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