Cryoradiolysis of oxygenated cytochrome P450 17A1 with lyase substrates generates expected products.
J Inorg Biochem
; 257: 112582, 2024 Aug.
Article
em En
| MEDLINE
| ID: mdl-38723329
ABSTRACT
When subjected to γ-irradiation at cryogenic temperatures the oxygenated complexes of Cytochrome P450 CYP17A1 (CYP17A1) bound with either of the lyase substrates, 17α-Hydroxypregnenolone (17-OH PREG) or 17α-Hydroxyprogesterone (17-OH PROG) are shown to generate the corresponding lyase products, dehydroepiandrosterone (DHEA) and androstenedione (AD) respectively. The current study uses gas chromatography-mass spectrometry (GC/MS) to document the presence of the initial substrates and products in extracts of the processed samples. A rapid and efficient method for the simultaneous determination of residual substrate and products by GC/MS is described without derivatization of the products. It is also shown that no lyase products were detected for similarly treated control samples containing no nanodisc associated CYP17 enzyme, demonstrating that the product is formed during the enzymatic reaction and not by GC/MS conditions, nor the conditions produced by the cryoradiolysis process.
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Base de dados:
MEDLINE
Assunto principal:
Esteroide 17-alfa-Hidroxilase
/
Cromatografia Gasosa-Espectrometria de Massas
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article