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Biochemistry and genetics of esterase-20 (ES-20), a second trimeric carboxylesterase of the house mouse (Mus musculus). I. Purification and characterization of ES-20C1 from male kidney

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Abstract

ES-201 was isolated from male mouse kidney and purified 350-fold by ion-exchange chromatography, isoelectric focusing, and gel filtration. The resultant product was apparently homogeneous by the criteria of polyacrylamide gel electrophoresis and immunodiffusion and represented a major fraction of male mouse kidney esterase. Sodium dodecyl sulfate gel electrophoresis revealed the presence of a single subunit band, molecular weight 59,500; the molecular weight of the native protein was found to be 179,000. Titration of the active site yielded an equivalent weight of about 175,000. The enzyme was further characterized by its kinetic parameters for the hydrolysis of a series of 4-nitrophenyl esters and was classified as a carboxylesterase (EC 3.1.1.1). ES-20C1 bound to concanavalin A, indicating that it was a high-mannose-type glycoprotein; the role of terminal β-N-acetylglucosamine residues in the carbohydrate side chains for stabilization of the quaternary structure of the trimer was revealed. Extensive biochemical and immunological similarities to ES-9C supported an earlier suggestion that theEs-9 c gene product is a component of the ES-20C1 trimer.

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Authors and Affiliations

  1. Abteilung für Chemische Pathologie, Pathologisches Institut der Universität Freiburg, Albertstrasse 19, D-7800, Freiburg i. Br., FRG

    S. M. de Looze, A. Ronai & O. H. von Deimling

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  1. S. M. de Looze
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  2. A. Ronai
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  3. O. H. von Deimling
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Additional information

This is communication No. 48 of a research program devoted to the genetics, cellular distribution, and function of nonspecific esterases. This work was supported by the Deutsche Forschungsgemeinschaft (De 315/2).

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de Looze, S.M., Ronai, A. & von Deimling, O.H. Biochemistry and genetics of esterase-20 (ES-20), a second trimeric carboxylesterase of the house mouse (Mus musculus). I. Purification and characterization of ES-20C1 from male kidney. Biochem Genet 23, 741–757 (1985). https://doi.org/10.1007/BF00554085

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  • DOI: https://doi.org/10.1007/BF00554085

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