氧合细胞色素 P450 17A1 的冷冻放射解作用与裂合底物生成预期产物。
Cryoradiolysis of oxygenated cytochrome P450 17A1 with lyase substrates generates expected products.
机构信息
Department of Chemistry, Marquette University, P.O. Box 1881, Milwaukee, WI 53201-1881, USA; Department of Chemistry, University of Georgia, Athens, GA 30602, USA.
Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
出版信息
J Inorg Biochem. 2024 Aug;257:112582. doi: 10.1016/j.jinorgbio.2024.112582. Epub 2024 May 4.
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.
当在低温下经受γ辐照时，与裂解酶底物 17α-羟孕烯醇酮（17-OH PREG）或 17α-羟孕酮（17-OH PROG）结合的细胞色素 P450 CYP17A1（CYP17A1）的含氧复合物分别生成相应的裂解酶产物脱氢表雄酮（DHEA）和雄烯二酮（AD）。本研究使用气相色谱-质谱联用（GC/MS）来记录处理样品提取物中初始底物和产物的存在。描述了一种无需产物衍生化即可同时测定残留底物和产物的快速高效 GC/MS 方法。还表明，对于含有与纳米盘相关的 CYP17 酶的对照样品，没有检测到裂解酶产物，这表明产物是在酶反应过程中形成的，而不是由 GC/MS 条件、也不是由低温放射分解过程产生的条件形成的。
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