The estimation of dihydropteridine reductase in human blood cells.
Significant homology between dihydropteridine reductase (DHPR) from rat and human sources has been established by the ability of polyclonal antibodies raised to the rat-liver enzyme to detect the human protein in Western blots. The antibody also reacted with a single protein in bovine, dog and porcine kidney extracts, however, only trace reactivity was detected in rabbit. Quantitation of Western blots by soft laser densitometry showed that the response was proportional to total protein present in analyses of both pure rat-liver enzyme samples and crude extracts of rat and human liver. The DHPR contents of human blood cells were analysed by this method and the results compared to levels determined in enzymatic assays. Extracts of platelets and lymphocytes showed good correlation between these two methods, however, granulocytes exhibited high apparent enzyme activity but no DHPR protein detectable in blots. Erythrocyte extracts showed approximately 50% lower DHPR protein levels than predicted by activity measurements. These results are discussed in relation to the accuracy of detecting DHPR deficiencies in humans by enzymatic assay of whole blood samples.
Webber
S
S
Department of Basic and Clinical Research, Research Institute of Scripps Clinic, La Jolla, CA.
Hural
J A
JA
Whiteley
J M
JM
D000818:Animals; D000906:Antibodies; D001773:Blood Cells / Q000201:enzymology*; D004093:Dihydropteridine Reductase / Q000097:blood*; D004586:Electrophoresis; D006801:Humans; D008099:Liver / Q000201:enzymology; D009247:NADH, NADPH Oxidoreductases / Q000097:blood*; D011817:Rabbits; D051381:Rats
D000906:Antibodies; D004093:Dihydropteridine Reductase; D009247:NADH, NADPH Oxidoreductases