Synthesis of a putative substrate for malonyl-coenzyme A: 21-hydroxypregnane 21-O-malonyltransferase and development of an HPLC method for the quantification of the enzyme reaction
| Autor: | Wolfgang Kreis, Serge Philibert Kuate, Herve M. P. Poumale, Rodrigo Maia de Pádua |
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| Rok vydání: | 2008 |
| Předmět: |
Chromatography
Gas Coenzyme A Clinical Biochemistry 01 natural sciences Biochemistry Chemical synthesis Sensitivity and Specificity Analytical Chemistry Substrate Specificity chemistry.chemical_compound Cardenolide Side chain Chromatography High Pressure Liquid Butenolide Chromatography 010405 organic chemistry Substrate (chemistry) Reproducibility of Results Cell Biology General Medicine Malonyl Coenzyme A 0104 chemical sciences 010404 medicinal & biomolecular chemistry chemistry Spectrophotometry Ultraviolet Chromatography Thin Layer Derivative (chemistry) Acyltransferases |
| Zdroj: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 860(2) |
| ISSN: | 1570-0232 |
| Popis: | The butenolide ring is the main common characteristic of all cardenolides. Its formation is supposed to be initiated by the transfer of a malonyl moiety from malonyl-coenzyme A to an appropriate 21-hydroxypregnane. A new, reliable, fast and sensitive method to determine malonyl-coenzyme A: 21-hydroxypregnane 21-O-malonyltransferase activity had to be developed since previous attempts employing HPLC, TLC or GC did not prove successful. A surrogate substrate was synthesized containing a side chain resembling the sugar side chain attached to C-3 of putative cardenolide precursors and containing a chromophor allowing UV detection. 3beta-benzoyloxy-5beta-pregnane-14beta,21-dihydroxy-20-one and its 21-O-malonylated derivative were synthesized, the latter being the expected product of the enzyme reaction. The new substrate was well accepted by the enzyme. An HPLC method has been established to detect and quantify 3beta-benzoyloxy-5beta-pregnane-14beta,21-dihydroxy-20-one and its 21-O-malonylated derivative, 3beta-benzoyloxy-5beta-pregnane-14beta-hydroxy-20-one 21-O-malonylhemiester. The method was validated. |
| Databáze: | OpenAIRE |
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