PMID: 2510599Nov 15, 1989Paper

Purification of tyrosinase to homogeneity based on its resistance to sodium dodecyl sulfate-proteinase K digestion

Archives of Biochemistry and Biophysics
E J Yurkow, J D Laskin

Abstract

A two-step method was used to purify enzymatically active tyrosinase to apparent homogeneity from B16/C3 mouse melanoma cells. The purification was based on our finding that tyrosinase was highly resistant to sodium dodecyl sulfate-proteinase K digestion. Following treatment with proteinase K, tyrosinase was separated from all detectable contaminants using DEAE-52 ion-exchange chromatography. Using this procedure, tyrosinase was purified to high specific activity and in yields greater than 50%. The purified enzyme was then used to generate high titers of specific polyclonal antibodies in rabbits. These antibodies were used to immunoprecipitate all tyrosinase isozymes from murine melanoma tumor extracts and cultured B16/C3 cells metabolically labeled with [35S]methionine.

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Citations

Jul 15, 1991·Experientia·A Slominski, R Costantino
Sep 1, 1991·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·A Slominski
Aug 6, 1991·Biochimica Et Biophysica Acta·J M Wood, K U Schallreuter
Oct 27, 1998·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·S HataH Yokosawa
Sep 6, 2014·Molecular Biotechnology·Stefano FogalElisabetta Bergantino
Sep 1, 1992·Archives of Biochemistry and Biophysics·P ValverdeJ A Lozano
Mar 3, 2018·International Journal of Molecular Sciences·Tobias MannLudger Kolbe
May 14, 2017·Scientific Reports·Matthias PretzlerAnnette Rompel
Dec 8, 1998·Phytochemistry·S MariJ J Macheix
Apr 11, 2009·Molecular Pharmaceutics·Luís Sánchez-del-CampoJosé Neptuno Rodríguez-López
Dec 1, 1991·The Journal of Investigative Dermatology·K U SchallreuterJ Berger
May 1, 1996·The Journal of Investigative Dermatology·S ChenM A Philbert
Feb 1, 1991·The Journal of Investigative Dermatology·A SlominskiR Costantino
Oct 28, 2008·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·I MaidaR Cicero

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