Determination of microsomal lauric acid hydroxylase activity by HPLC with flow-through radiochemical quantitation
Determination of microsomal lauric acid hydroxylase activity by HPLC with flow-through radiochemical quantitation
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HPLC 流通式放射化学定量法测定微粒体月桂酸羟化酶活性
DOI：
10.1016/0003-2697(88)90093-0
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发表时间：
1988
影响因子：
2.9
通讯作者：
J. Newton
中科院分区：
文献类型：
--
作者：
M. Romano;K. Straub;L. Yodis;Regina D. Eckardt;J. Newton
本文建立了一种基于高效液相色谱-流通放射化学检测的微粒体月桂酸羟基化测定方法。优化了LA的三种微粒体代谢物的分离度和定量条件，其中一种以前未报告为哺乳动物微粒体孵育中的LA代谢物。这些产物，12-（ω）-羟基-LA，11-（ω-1）-羟基-LA，和一个新的代谢产物，10-（ω-2）-羟基-LA的分离和气相色谱/质谱鉴定。在NADPH存在下，所有三种代谢产物的形成与时间和微粒体蛋白浓度呈线性关系。过氧化氢也支持LA的微粒体代谢，尽管代谢物的比例与NADPH支持的微粒体产生的代谢物的比例有很大不同。用几种生化探针（甲吡酮、α-萘酮、2-二乙基氨基乙基-2，2-二苯基戊酸酯盐酸盐和10-十一炔酸）解离这三种LA羟化酶活性。这些实验表明，LA的位点特异性羟基化[ω-，（ω-1）-，（ω-2）-]可能由细胞色素P-450的不同同工酶催化。
An assay for the microsomal hydroxylation of lauric acid (LA), based on HPLC with flow-through radiochemical detection, has been developed. Conditions were optimized for resolution and quantitation of three microsomal metabolites of LA, one of which has not been reported previously as a metabolite of LA in mammalian microsomal incubations. These products, 12-(ω)-hydroxy-LA, 11-(ω-1)-hydroxy-LA, and a novel metabolite, 10-(ω-2)-hydroxy-LA, were isolated by HPLC and identified by gas chromatography/mass spectrometry. In the presence of NADPH, the formation of all three metabolites was linear with time and microsomal protein concentration. Hydrogen peroxide also supported the microsomal metabolism of LA, although the ratio of metabolites was substantially different than that produced by NADPH-supported microsomes. Several biochemical probes (metyrapone, α-naphthoflavone, 2-diethylaminoethyl-2,2-diphenylvalerate hydrochloride, and 10-undecynoic acid) were used to dissociate the three LA hydroxylase activities. These experiments suggest that the site-specific hydroxylation [ω-, (ω-1)-, (ω-2)-] of LA may be catalyzed by different isozymes of cytochrome P-450.
乙炔脂肪酸对肝脂肪酸羟化酶的特异性灭活。
DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
OrtizdeMontellano,PR;Reich,NO
通讯作者：
Reich,NO
在甲基胆蒽治疗和对照大鼠中，NADPH 与 H2O2 支持的肝微粒体对前列腺素羟基化的区域选择性：新型前列腺素代谢物的形成。
影响因子：
3.9
作者：
Holm,KA;Engell,RJ;Kupfer,D
通讯作者：
Kupfer,D
猪肾微粒体和纯化的肾细胞色素 P-450 对前列腺素 A1 和 E1 以及月桂酸的 omega- 和 (omega-1)-羟化酶活性。
DOI：
--
发表时间：
1981
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Okita,RT;Parkhill,LK;Yasukochi,Y;Masters,BS;Theoharides,AD;Kupfer,D
通讯作者：
Kupfer,D