The determination of homocysteine-thiolactone in biological samples

被引：86

作者：

Jakubowski, H ^{[1
]}

机构：

[1] Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Microbiol & Mol Genet, Int Ctr Publ Hlth, Newark, NJ 07101 USA

[2] Polish Acad Sci, Inst Bioorgan Chem, PL-61704 Poznan, Poland

来源：

ANALYTICAL BIOCHEMISTRY | 2002年 / 308卷 / 01期

基金：

美国国家科学基金会;

关键词：

homocysteine-thiolactone; HPLC; human endothelial cell culture; human plasma; yeast; bacteria;

D O I：

10.1016/S0003-2697(02)00224-5

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Homocysteine-thiolactone, a cyclic thioester of homocysteine, is synthesized by methionyl-tRNA synthetase in all cell types. A new assay for the determination of homocysteine-thiolactone in biological samples is described. The assay involves separation of homocysteine-thiolactone from macromolecules by ultrafiltration. Homocysteine-thiolactone is further purified and quantified by high-pressure liquid chromatography either on a reverse phase or a cation exchange micro-bore column. The detection and quantitation are obtained by monitoring the absorbance at 240 nm, a maximum in a UV spectrum of homocysteine-thiolactone. The sensitivity of detection is 5 pmol. This assay has been applied to bacteria (Escherichia coli and Mycobacterium smegmatis), the yeast Saccharomyces cerevisiae, cultured human vascular endothelial cells, and human plasma. The data support the conclusion that homocysteine-thiolactone is a ubiquitous metabolite whose levels are directly related to homocysteine levels. (C) 2002 Elsevier Science (USA). All rights reserved.

引用

页码：112 / 119

页数：8

共 30 条

[1] RADIOLYSIS OF AQUEOUS-SOLUTIONS OF HOMOCYSTEINETHIOLACTONE HYDROCHLORIDE [J].

ANDERSON, RF ;

PACKER, JE .

INTERNATIONAL JOURNAL FOR RADIATION PHYSICS AND CHEMISTRY, 1974, 6 (01) :33-46

[2]

Baernstein HD, 1934, J BIOL CHEM, V106, P451

[3] ROLE OF CARBOXY-TERMINAL REGION IN PROOFREADING FUNCTION OF METHIONYL-TRANSFER-RNA SYNTHETASE IN ESCHERICHIA-COLI [J].

GAO, WW ;

GOLDMAN, E ;

JAKUBOWSKI, H .

BIOCHEMISTRY, 1994, 33 (38) :11528-11535

[4] Homocysteine thiolactone is a positive effector of σS levels in Escherichia coli [J].

Goodrich-Blair, H ;

Kolter, R .

FEMS MICROBIOLOGY LETTERS, 2000, 185 (02) :117-121

[5] CHARACTERIZATION OF A HUMAN KAPOSIS-SARCOMA CELL-LINE THAT INDUCES ANGIOGENIC TUMORS IN ANIMALS [J].

HERNDIER, BG ;

WERNER, A ;

ARNSTEIN, P ;

ABBEY, NW ;

DEMARTIS, F ;

COHEN, RL ;

SHUMAN, MA ;

LEVY, JA .

AIDS, 1994, 8 (05) :575-581

[6] SYNTHESIS OF HOMOCYSTEINE THIOLACTONE BY METHIONYL-TRANSFER RNA-SYNTHETASE IN CULTURED-MAMMALIAN-CELLS [J].

JAKUBOWSKI, H ;

GOLDMAN, E .

FEBS LETTERS, 1993, 317 (03) :237-240

[7] Calcium-dependent human serum homocysteine thiolactone hydrolase -: A protective mechanism against protein N-homocysteinylation [J].

Jakubowski, H .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (06) :3957-3962

[8] Homocysteine thiolactone and protein homocysteinylation in human endothelial cells - Implications for atherosclerosis [J].

Jakubowski, H ;

Zhang, L ;

Bardeguez, A ;

Aviv, A .

CIRCULATION RESEARCH, 2000, 87 (01) :45-51

[9]

Jakubowski H, 1997, J BIOL CHEM, V272, P1935

[10] Aminoacylation of coenzyme A and pantetheine by aminoacyl-tRNA synthetases: Possible link between noncoded and coded peptide synthesis [J].

Jakubowski, H .

BIOCHEMISTRY, 1998, 37 (15) :5147-5153

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