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Lymphocyte Phosphatase-Associated Phosphoprotein Is a Substrate of Protein Kinase CK2

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Abstract

Lymphocyte phosphatase-associated phosphoprotein (LPAP) is a molecular partner of CD45 phosphatase that plays a key role in the regulation of antigen-specific activation of lymphocytes. The functions of LPAP still remain unknown. We believe that studying LPAP phosphorylation pathways could shed light on its functions. In this work, we studied the phosphorylation of LPAP ectopically expressed in non-lymphoid cells in order to determine the effect of LPAP interaction partners on its phosphorylation. We found that phosphorylation at Ser153 and Ser163 in non-hematopoietic HEK293 cells was conserved, while phosphorylation at Ser99 and Ser172 was almost absent. The pattern of LPAP phosphorylation in K562 erythroid and U937 myeloid cells expressing endogenous CD45 protein was similar to that observed in T and B lymphocytes. We demonstrated for the first time that LPAP is a substrate for protein kinase CK2 that phosphorylates it at Ser153, presumably ensuring LPAP resistance to degradation.

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Abbreviations

CIP:

calf intestinal phosphatase

CK2:

casein kinase 2

2D-DIGE:

two-dimensional difference gel electrophoresis

DTT:

dithiothreitol

IEF:

isoelectric focusing

LPAP:

lymphocyte phosphatase-associated phosphoprotein

PAGE:

polyacrylamide gel electrophoresis

PMA:

phorbol 12-myristate 13-acetate

SDS:

sodium dodecyl sulfate

TBB:

4,5,6,7-tetrabromo-2-azabenzimidazole

WB:

Western blotting

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Authors and Affiliations

  1. Institute of Immunology National Research Center, Federal Medical-Biological Agency, 115522, Moscow, Russia

    T. D. Tsoy, N. A. Kruglova & A. V. Filatov

  2. Faculty of Biology, Lomonosov Moscow State University, 119234, Moscow, Russia

    T. D. Tsoy, N. A. Kruglova & A. V. Filatov

Authors
  1. T. D. Tsoy
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  2. N. A. Kruglova
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  3. A. V. Filatov
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Corresponding author

Correspondence to A. V. Filatov.

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Published in Russian in Biokhimiya, 2018, Vol. 83, No. 11, pp. 1698–1707.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM18-184, October 22, 2018.

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Tsoy, T.D., Kruglova, N.A. & Filatov, A.V. Lymphocyte Phosphatase-Associated Phosphoprotein Is a Substrate of Protein Kinase CK2. Biochemistry Moscow 83, 1380–1387 (2018). https://doi.org/10.1134/S0006297918110081

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  • DOI: https://doi.org/10.1134/S0006297918110081

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