A differential microdetermination for the various forms of biopterin. |
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Authors: | T Fukushima K Kobayashi I Eto T Shiota |
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Affiliation: | 1. Department of Biology, Tokyo Metropolitan University, Setagaya, Tokyo 158, Japan;2. Department of Microbiology, The University of Alabama in Birmingham, University Station, Birmingham, Alabama 35294 USA;3. The Institute of Dental Research, The University of Alabama in Birmingham, University Station, Birmingham, Alabama 35294 USA |
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Abstract: | Conditions for the quantitative oxidation and destruction of tetrahydrobiopterin and quinoid dihydrobiopterin and the separation of biopterin from its reduced forms by ECTEOLA-Sephadex column chromatography are described. A procedure for the quantitation of tetrahydrobiopterin plus quinoid dihydrobiopterin, 7,8-dihydrobiopterin, and biopterin using a Crithidia bioassay is presented. Using these procedures it was found that tetrahydrobiopterin plus quinoid dihydrobiopterin are the prevalent forms in liver and blood of mice and that biopterin was the predominant form in the tails of tadpoles. In human urine, approximately half of the biopterin was found as tetrahydrobiopterin plus quinoid dihydrobiopterin and the other half was 7,8-dihydrobiopterin. The presence of tetrahydrobiopterin and quinoid dihydrobiopterin was confirmed by a coenzyme assay for the hydroxylation of phenylalanine. |
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