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A protein tyrosine kinase associated with the ATP-dependent inactivation of adipose diacylglycerol acyltransferase

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Lipids

Abstract

The activity that has been previously reported to reversibly inactivate adipose glycerolphosphate acyltransferase (GPAT) and diacylglycerol acyltransferase (DGAT)in vitro in the presence of ATP is shown here to be partially purified from adipose tissue with an apparent molecular weight of 68 kDa. The activity responsible for inactivating DGAT is associated with a kinase activity as determined by phosphate incorporation both into microsomal proteins and into a synthetic tyrosine-containing peptide as substrate for protein tyrosine kinase. Two microsomal polypeptides of 53 and 69 kDa are major substrates of this kinase. Both DGAT inactivating and kinase activities assayed from the purified sample have been found to be insensitive to the Ser/Thr kinase inhibitor H-7 while being sensitive to genistein and tyrphostin-25. A crude protein phosphatase preparation from liver was capable of reversing the effects of both activities. The purified sample was also shown to inactivate GPAT in the presence of ATP. These results suggest that a protein tyrosine kinase, in concert with a protein tyrosine phosphatase, may regulate the activities of DGAT and GPAT by a phosphorylation-dephosphorylation mechanism.

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Abbreviations

BSA:

bovine serum albumin

DGAT:

diacylglycerol acyltransferase

DTT:

dl-dithiothreitol

GPAT:

glycerolphosphate acyltransferase

PAGE:

polyacrylamine gel electrophoresis

PAP:

phosphatidate phosphatase

PKA:

protein kinase A

PKC:

protein kinase C

TAG:

triacylglycerol

TED:

buffer, 50 mM Tris-HCl (pH 7.5)/l mM EDTA/2 mM DTT

TMD:

buffer, 50 mM Tris-HCl (pH 7.5)/10 mM MgCl2/1 mM DTT

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Authors and Affiliations

  1. Department of Biochemistry, University of Ottawa, 365 Nicholas, P.O. Box 450 Stn. A, K1N 6N5, Ottawa, Ontario, Canada

    T. E. Lau & M. A. Rodriguez

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  1. T. E. Lau
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  2. M. A. Rodriguez
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Lau, T.E., Rodriguez, M.A. A protein tyrosine kinase associated with the ATP-dependent inactivation of adipose diacylglycerol acyltransferase. Lipids 31, 277–283 (1996). https://doi.org/10.1007/BF02529874

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  • DOI: https://doi.org/10.1007/BF02529874

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