PURIFICATION AND CHARACTERIZATION OF CARBOXYLESTERASES FROM RAT LUNG

被引:24
作者
GAUSTAD, R [1 ]
SLETTEN, K [1 ]
LOVHAUG, D [1 ]
FONNUM, F [1 ]
机构
[1] UNIV OSLO,DEPT BIOCHEM,N-0316 OSLO 3,NORWAY
关键词
D O I
10.1042/bj2740693
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
Carboxylesterase (EC 3.1.1.1) has played an important part in our understanding of the toxicokinetic behaviour of the organophosphorus cholinesterase inhibitors. Carboxylesterases are a heterogeneous group of enzymes that can be separated on the basis of their isoelectric points and by their substrate-specificity. We have purified the isoenzyme (pI 5.8) present in greatest activity in rat lung to near homogeneity. The enzyme was purified by (NH4)2SO4 precipitation, gel filtration, chromatofocusing, separation on anion- and cation-exchangers and hydrophobic-interaction chromatography. The purified enzyme has a molecular mass of approx. 180 kDa with subunits of 60 kDa. The substrate and inhibitor specificities of the enzyme have been characterized. Edman degradation revealed the first 19 amino acid residues of the enzyme. The N-terminus was found to be tyrosine. Inhibition of the enzyme by organophosphorus compounds differed greatly from that of cholinesterases, despite the partial analogy at the N-terminal region.
引用
收藏
页码:693 / 697
页数:5
相关论文
共 26 条
  • [1] ALDRIDGE WN, 1972, ENZYME INHIBITORS SU
  • [2] ALI B, 1985, P W PHARMACOL SOC, V28, P319
  • [3] THE PREPARATION AND PHYSICAL PROPERTIES OF ISOPROPYL METHYLPHOSPHONOFLUORIDATE (SARIN)
    BRYANT, PJR
    FORDMOORE, AH
    PERRY, BJ
    WARDROP, AWH
    WATKINS, TF
    [J]. JOURNAL OF THE CHEMICAL SOCIETY, 1960, (APR): : 1553 - 1555
  • [4] EVIDENCE THAT THE AMYLOID FIBRIL PROTEIN IN SENILE SYSTEMIC AMYLOIDOSIS IS DERIVED FROM NORMAL PREALBUMIN
    CORNWELL, GG
    SLETTEN, K
    JOHANSSON, B
    WESTERMARK, P
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1988, 154 (02) : 648 - 653
  • [5] CARBOXYLESTERASES, IMPORTANCE FOR DETOXIFICATION OF ORGANOPHOSPHORUS ANTICHOLINESTERASES AND TRICHOTHECENES
    FONNUM, F
    STERRI, SH
    AAS, P
    JOHNSEN, H
    [J]. FUNDAMENTAL AND APPLIED TOXICOLOGY, 1985, 5 (06): : S29 - S38
  • [6] GOEPPINGER A, 1978, Biochimica et Biophysica Acta, V525, P74
  • [7] Heymann E, 1981, Methods Enzymol, V77, P405
  • [8] CHARACTERIZATION OF THE ISOENZYMES OF PIG-LIVER ESTERASE .1. CHEMICAL STUDIES
    HEYMANN, E
    JUNGE, W
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1979, 95 (03): : 509 - 518
  • [9] INVESTIGATION OF MICROHETEROGENEITY AND SUBUNIT STRUCTURE OF CARBOXYLESTERASES BY QUANTITATIVE-DETERMINATION OF TERMINAL AMINO-ACIDS
    HEYMANN, E
    IGLESIA, J
    [J]. HOPPE-SEYLERS ZEITSCHRIFT FUR PHYSIOLOGISCHE CHEMIE, 1974, 355 (09): : 1112 - 1120
  • [10] STRUCTURE OF ACTIVE CENTRE OF MICROSOMAL CARBOXYL ESTERASES FROM PIG KIDNEY AND LIVER
    HEYMANN, E
    KRISCH, K
    PAHLICH, E
    [J]. HOPPE-SEYLERS ZEITSCHRIFT FUR PHYSIOLOGISCHE CHEMIE, 1970, 351 (08): : 931 - &