PURIFICATION OF NEUROPATHY TARGET ESTERASE FROM AVIAN BRAIN AFTER PRELABELING WITH [H-3] DIISOPROPYL PHOSPHOROFLUORIDATE

被引：15

作者：

RUFFERTURNER, ME

READ, DJ

JOHNSON, MK

机构：

[1] Mrc Toxicology Unit, Medical Research Council Laboratories, Surrey

来源：

JOURNAL OF NEUROCHEMISTRY | 1992年 / 58卷 / 01期

关键词：

NEUROPATHY TARGET ESTERASE; H-3]DIISOPROPYL PHOSPHOROFLUORIDATE LABELING; PURIFICATION;

D O I：

10.1111/j.1471-4159.1992.tb09288.x

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：

Neuropathy target esterase from hen brains was radiolabelled at the active site with [H-3]diisopropyl phosphorofluoridate. The labelled protein was purified by differential centrifugation and Nonidet P40 solubilization, detergent phase partitioning, anion exchange, and preparative sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The volatilizable counts assay and analytical SDS-PAGE were used to monitor the protein. The 150-kDa subunit polypeptide appears as a single band on analytical SDS-PAGE.

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页码：135 / 141

页数：7

相关论文

共 26 条

[1]

BORDIER C, 1981, J BIOL CHEM, V256, P1604

[2] A VERSATILE SYSTEM FOR PREPARATIVE ELECTROPHORESIS IN ACRYLAMIDE GEL [J].

BROWNSTO.AD .

ANALYTICAL BIOCHEMISTRY, 1969, 27 (01) :25-&

[3] FURTHER DEVELOPMENT OF A VERSATILE SYSTEM FOR PREPARATIVE ELECTROPHORESIS IN ACRYLAMIDE-GEL [J].

BROWNSTONE, AD .

ANALYTICAL BIOCHEMISTRY, 1976, 70 (02) :572-591

[4] CHARACTERIZATION OF [3H]DI-ISOPROPYL PHOSPHOROFLUORIDATE-BINDING PROTEINS IN HEN BRAIN - RATES OF PHOSPHORYLATION AND SENSITIVITY TO NEUROTOXIC AND NON-NEUROTOXIC ORGANO-PHOSPHORUS COMPOUNDS [J].

CARRINGTON, CD ;

ABOUDONIA, MB .

BIOCHEMICAL JOURNAL, 1985, 228 (03) :537-544

[5] TARGET SIZE OF NEUROTOXIC ESTERASE AND ACETYLCHOLINESTERASE AS DETERMINED BY RADIATION INACTIVATION [J].

CARRINGTON, CD ;

FLUKE, DJ ;

ABOUDONIA, MB .

BIOCHEMICAL JOURNAL, 1985, 231 (03) :789-792

[6] NEUROTOXIC ESTERASE - GEL-FILTRATION AND ISOELECTRIC-FOCUSING OF CARBOXYLESTERASES SOLUBILIZED FROM HEN BRAIN [J].

CHEMNITIUS, JM ;

HASELMEYER, KH ;

ZECH, R .

LIFE SCIENCES, 1984, 34 (12) :1119-1125

[7] RAPID AGING OF NEUROTOXIC ESTERASE AFTER INHIBITION BY DI-ISOPROPYL PHOSPHOROFLUORIDATE [J].

CLOTHIER, B ;

JOHNSON, MK .

BIOCHEMICAL JOURNAL, 1979, 177 (02) :549-558

[8] NEUROTOXIC ESTERASE - CHARACTERIZATION OF THE SOLUBILIZED ENZYME AND THE CONDITIONS FOR ITS SOLUBILIZATION FROM CHICKEN BRAIN MICROSOMAL-MEMBRANES WITH IONIC, ZWITTERIONIC, OR NONIONIC DETERGENTS [J].

DAVIS, CS ;

RICHARDSON, RJ .

BIOCHEMICAL PHARMACOLOGY, 1987, 36 (09) :1393-1399

[9]

JOHNSON MK, 1971, BIOCHEM J, V122, pP51

[10] DELAYED NEUROTOXIC EFFECT OF SOME ORGANOPHOSPHORUS COMPOUNDS - IDENTIFICATION OF PHOSPHORYLATION SITE AS AN ESTERASE [J].

JOHNSON, MK .

BIOCHEMICAL JOURNAL, 1969, 114 (04) :711-&