HPLC-MS/MS analysis of the products generated from all-trans-retinoic acid using recombinant human CYP26A

被引：65

作者：

Chithalen, JV

Luu, L

Petkovich, M

Jones, G ^{[1
]}

机构：

[1] Queens Univ, Dept Biochem, Kingston, ON K7L 3N6, Canada

[2] Queens Univ, Dept Pathol, Kingston, ON K7L 3N6, Canada

[3] Queens Univ, Dept Med, Kingston, ON K7L 3N6, Canada

来源：

JOURNAL OF LIPID RESEARCH | 2002年 / 43卷 / 07期

关键词：

vitamin A metabolism; retinoids; cytochrome P450; RAI-1;

D O I：

10.1194/jlr.M100343-JLR200

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Two mammalian hCYP26A expression systems have been used to analyze the metabolic products of CYP26A. Through the use of extensive HPLC, UV spectroscopy, and liquid chromatography/tandem mass spectrometry (LC-MS/MS) methodology, we have conclusively demonstrated that the complex mixture of products comprises 4-OH-all-trans-retinoic acid, 4-oxo-all-trans-retinoic acid, and 18-OH-all-trans-retinoic acid, and more polar products, partially identified as dihydroxy and mono-oxo, mono-hydroxy derivatives. These more polar products are presumed to result from multiple hydroxylations on the beta-ionone ring. The inter-relationship of initial and polar metabolites was inferred from both gene-dose and time-course experiments. Both initial and secondary metabolic steps after 4-oxo-all-trans-retinoic acid are ketoconazole-sensitive, suggesting that steps in the production of water-soluble metabolites are cytochrome P450-dependent. HPLC-MS/MS analysis of the products generated from all-trans-retinoic acid using recombinant human CYP26A.

引用

页码：1133 / 1142

页数：10

共 43 条

[1] The retinoic acid-metabolizing enzyme, CYP26A1, is essential for normal hindbrain patterning, vertebral identity, and development of posterior structures [J].