ASSAY FOR ENKEPHALIN-GENERATING ENZYME IN RAT-BRAIN TISSUES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH POSTCOLUMN FLUORESCENCE DERIVATIZATION

被引：4
作者：
ZHANG, GQ
KAI, M
OHKURA, Y
机构：
[1] Faculty of Pharmaceutical Sciences, Kyushu University 62, Maidashi
来源：
ANALYTICAL SCIENCES | 1991年 / 7卷 / 04期
关键词：
ENZYME ASSAY; ENKEPHALIN-GENERATING ENZYME; RAT BRAIN; HIGH PERFORMANCE LIQUID CHROMATOGRAPHY; POSTCOLUMN DERIVATIZATION; OPIOID PEPTIDE;
D O I：
10.2116/analsci.7.561
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
A simple and sensitive method for assaying enkephalin-generating enzyme activity in rat brain tissues is described. Methionine-enkephalin, produced enzymatically from alpha-endorphin and BAM-12P as substrates and leucine-enkephalin from alpha-neoendorphin, were quantified by high performance liquid chromatography with postcolumn fluorescence derivatization using hydroxylamine, cobalt(II) and borate. The detection limits for methionine-and leucine-enkephalin-generating activities of the enzyme are 2.5 and 3.0 pmol min-1 mg-protein-1, respectively. The enzyme can be characterized as an endo-oligopeptidase A-like enzyme on the basis of inhibition studies.
引用
收藏
页码：561 / 565
页数：5
相关论文
共 19 条
[1]   PREPARATION, ASSAY, AND PARTIAL CHARACTERIZATION OF A NEUTRAL ENDOPEPTIDASE FROM RABBIT BRAIN [J].
CAMARGO, ACM ;
SHAPANKA, R ;
GREENE, LJ .
BIOCHEMISTRY, 1973, 12 (09) :1838-1844
[2]   CONVERSION AND INACTIVATION OF OPIOID-PEPTIDES BY RABBIT BRAIN ENDO-OLIGOPEPTIDASE-A [J].
CAMARGO, ACM ;
RIBEIRO, MJVF ;
SCHWARTZ, WN .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1985, 130 (02) :932-938
[3]   BRAIN ENDO-OLIGOPEPTIDASE-A, A PUTATIVE ENKEPHALIN CONVERTING ENZYME [J].
CAMARGO, ACM ;
OLIVEIRA, EB ;
TOFFOLETTO, O ;
METTERS, KM ;
ROSSIER, J .
JOURNAL OF NEUROCHEMISTRY, 1987, 48 (04) :1258-1263
[4]   PURIFICATION OF RABBIT BRAIN ENDOOLIGOPEPTIDASES AND PREPARATION OF ANTI-ENZYME ANTIBODIES [J].
CARVALHO, KM ;
CAMARGO, ACM .
BIOCHEMISTRY, 1981, 20 (25) :7082-7088
[5]  
FRICKER LD, 1983, J BIOL CHEM, V258, P950
[6]   ENKEPHALIN CONVERTASE - PURIFICATION AND CHARACTERIZATION OF A SPECIFIC ENKEPHALIN-SYNTHESIZING CARBOXYPEPTIDASE LOCALIZED TO ADRENAL CHROMAFFIN GRANULES [J].
FRICKER, LD ;
SNYDER, SH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1982, 79 (12) :3886-3890
[7]  
HOOK VYH, 1985, J BIOL CHEM, V260, P5991
[8]   IDENTIFICATION OF 2 RELATED PENTAPEPTIDES FROM BRAIN WITH POTENT OPIATE AGONIST ACTIVITY [J].
HUGHES, J ;
SMITH, TW ;
KOSTERLITZ, HW ;
FOTHERGILL, LA ;
MORGAN, BA ;
MORRIS, HR .
NATURE, 1975, 258 (5536) :577-579
[9]   ISOLATION OF AN ENDOGENOUS COMPOUND FROM BRAIN WITH PHARMACOLOGICAL PROPERTIES SIMILAR TO MORPHINE [J].
HUGHES, J .
BRAIN RESEARCH, 1975, 88 (02) :295-308
[10]   SELECTIVE DETERMINATION OF N-TERMINAL TYROSINE-CONTAINING PEPTIDES BY A NOVEL FLUORESCENCE REACTION WITH BORATE, HYDROXYLAMINE AND COBALT(II) [J].
KAI, M ;
OHKURA, Y .
ANALYTICA CHIMICA ACTA, 1986, 182 :177-183
12