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
[11]   ALPHA-NEO-ENDORPHIN - BIG LEU-ENKEPHALIN WITH POTENT OPIATE ACTIVITY FROM PORCINE HYPOTHALAMI [J].
KANGAWA, K ;
MATSUO, H ;
IGARASHI, M .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1979, 86 (01) :153-160
[12]  
KNIGHT M, 1978, J BIOL CHEM, V253, P3843
[13]   FURTHER CHARACTERIZATION OF AN ENKEPHALIN-GENERATING ENZYME FROM ADRENAL-MEDULLARY CHROMAFFIN GRANULES [J].
LINDBERG, I ;
YANG, HYT ;
COSTA, E .
JOURNAL OF NEUROCHEMISTRY, 1984, 42 (05) :1411-1419
[14]  
LOWRY OH, 1951, J BIOL CHEM, V193, P265
[15]   MEMBRANE-BOUND TRYPSIN-LIKE-ENZYME FUNCTIONING IN DEGRADATION OF DYNORPHIN IN NEURO-BLASTOMA CELLS - PURIFICATION AND CHARACTERIZATION [J].
SATOH, M ;
YOKOSAWA, H ;
ISHII, S .
JOURNAL OF BIOCHEMISTRY, 1988, 103 (03) :493-498
[16]   ENKEPHALIN IS LIBERATED FROM METORPHAMIDE AND DYNORPHIN-A1-8 BY ENDO-OLIGOPEPTIDASE-A, BUT NOT BY METALLOENDOPEPTIDASE EC 3.4.24.15 [J].
TOFFOLETTO, O ;
METTERS, KM ;
OLIVEIRA, EB ;
CAMARGO, ACM ;
ROSSIER, J .
BIOCHEMICAL JOURNAL, 1988, 252 (01) :35-38
[17]   NEUROPEPTIDE PROCESSING ENZYMES [J].
TURNER, AJ .
TRENDS IN NEUROSCIENCES, 1984, 7 (07) :258-260
[18]   ENHANCEMENT OF DELAYED-TYPE HYPERSENSITIVITY BY BESTATIN, AN INHIBITOR OF AMINOPEPTIDASE-B AND LEUCINE AMINOPEPTIDASE [J].
UMEZAWA, H ;
ISHIZUKA, M ;
AOYAGI, T ;
TAKEUCHI, T .
JOURNAL OF ANTIBIOTICS, 1976, 29 (08) :857-859
[19]   DETERMINATION OF 7 OPIOID-PEPTIDES IN RAT-BRAIN BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH ONLINE POSTCOLUMN FLUORESCENCE DERIVATIZATION [J].
ZHANG, GQ ;
KAI, M ;
OHKURA, Y .
ANALYTICAL SCIENCES, 1990, 6 (05) :671-676
12