Cryoradiolysis of Oxygenated Cytochrome P450 17A1 with Lyase Substrates Generates Expected Products

Authors

• Remigio Usai, University of Georgia
• Ilia G. Denisov, University of Illinois at Urbana-Champaign
• Stephen G. Sligar, University of Illinois at Urbana-Champaign
• James R. Kincaid, Marquette UniversityFollow

Document Type

Article

Publication Date

8-2024

Publisher

Elsevier

Source Publication

Journal of Inorganic Biochemistry

Source ISSN

0162-0134

Original Item ID

DOI: 10.1016/j.jinorgbio.2024.112582

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.

Comments

Journal of Inorganic Biochemistry, Vol. 257 (August 2024). DOI.

Recommended Citation

Usai, Remigio; Denisov, Ilia G.; Sligar, Stephen G.; and Kincaid, James R., "Cryoradiolysis of Oxygenated Cytochrome P450 17A1 with Lyase Substrates Generates Expected Products" (2024). Chemistry Faculty Research and Publications. 1126.
https://epublications.marquette.edu/chem_fac/1126