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Protein tyrosine kinases in human breast cancer: Kinetic properties and evidence for the presence of two forms of native enzyme

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Summary

The protein tyrosine kinase (PTK) of human breast tumors classified as positive (TM +) or negative (TM -) according to their estrogen and progestin receptor levels was partially characterized with regard to its distribution, kinetic parameters, molecular size, and ability to phosphorylate endogenous mammary proteins. For both types of tumors, PTK activity depended upon the presence of Mn++ (2–5 mM) and/or Mg++ (10–20 mM). The activities, total (per g of tissue) and specific (per mg of protein), were similar for both types of tumors, and an average of 60% of activity was located in cytosolic fractions. The autoradiographic detection of alkali-resistant phosphoproteins after SDS-PAGE showed very similar patterns between corresponding fractions from both types of tumors. Upon gel filtration, two peaks of activity of apparent Mr 245 kDa (peak I) and 47 kDa (peak II) were observed. Peak II was found in both cytosols and extracts from particulate fractions, while peak I was present only in the latter fraction for both TM + and TM — tumors. The apparent Km's for ATP ranged from 4.1 to 6.6 µM, and from 11 to 34 µg/ml for the synthetic substrate poly [Glu80, Tyr20], at an optimal pH of 6.5–7.5. When endogenous alkali-resistant phosphorylation of peaks I and II was determined by autoradiography after SDS-PAGE, two major mammary proteins of Mr 60 and 45 kDa were phosphorylated by peak II and three, Mr 145, 74, and 62 kDa, by peak I. Thus, in TM + and TM — human breast cancer tissues, PTKs possess similar enzymatic and molecular properties. The Mr 47 kDa is present in both soluble and particulate fractions, while the Mr 245 kDa is only observed in the latter fraction.

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Authors and Affiliations

  1. Maisonneuve-Rosemont Research Center, 54151'Assomption Blvd., H1T 2M4, Montreal, Quebec, Canada

    Simone Chevalier

  2. Departments of Biochemistry and of Medicine, University of Montreal, Montreal, Quebec, Canada

    Yves Durocher

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  1. Yves Durocher
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  2. Simone Chevalier
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Durocher, Y., Chevalier, S. Protein tyrosine kinases in human breast cancer: Kinetic properties and evidence for the presence of two forms of native enzyme. Breast Cancer Res Tr 17, 99–107 (1990). https://doi.org/10.1007/BF01806290

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