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Phosphorylation of human interleukin-2 (IL-2)

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Abstract

Human interleukin-2 (IL-2) is a lymphokine which is capable of activating lymphocytes and supporting the long-term in vitro growth of activated T cell clones. Recombinant human IL-2, expressed in either E. coli or cos cells, was shown to be phosphorylated by protein kinase C. Phosphorylated IL-2 synthesized in E. coli was analyzed by SDS-PAGE, reverse phase HPLC, and tryptic peptide mapping. The phosphorylated tryptic peptide was identified as the N-terminal fragment containing a single phosphorylation site at the serine residue at position 7. There was no difference in biological activity between non-phosphorylated and phosphorylated IL-2, as determined by a T cell growth assay. Although the physiological role of phosphorylation of IL-2 is unclear, IL-2 can be labeled with [γ-32p] ATP and protein kinase C to a high specific radioactivity, and the synthesis of biologically active 32p-labeled IL-2 may be useful for receptor-binding studies of the cells containing low level of phosphoprotein phosphotases.

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Abbreviations

IL-2:

Interleukin-2

rIL-2:

recombinant IL-2

SDS-PAGE:

Sodium Dodecylsulfate Polyacrylamide Gel Electrophoresis, Tris tris (hydroxymethyl)-amino methane

RP-HPLC:

Reverse Phase High Pressure Liquid Chromatography

PTH:

Phenylthiohydantoin

IFN-α:

Leukocyte Interferon

IFN-β:

Fibroblast Interferon

IFN-γ:

Immune Interferon

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Authors and Affiliations

  1. LBP, BRMP, DCT NCI-FCRF Frederick, 21701, MD

    Hsiang-fu Kung & Ida Calvert

  2. Hoffman-LaRoche Inc., 07110, Nutley, NJ

    Eva Bekesi & Fazlur R Khan

  3. Endocrinology and Reproduction Research Branch, National Institute of Child Health, NIH, 20892, Bethesda, MD

    Kuo-ping Huang

  4. BRI CI-FCRF Frederick, 21701, MD

    Stephen Oroszlan, Louis E. Henderson, Terry D. Copeland & Raymond C. Sowder

  5. Department of Chemical Biology and Pharmacognosy, College of Pharmacy, Rutgers University, 08854, Piscataway, NJ, USA

    Shu-jing Wei

Authors
  1. Hsiang-fu Kung
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  2. Ida Calvert
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  3. Eva Bekesi
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  4. Fazlur R Khan
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  5. Kuo-ping Huang
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  6. Stephen Oroszlan
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  7. Louis E. Henderson
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  8. Terry D. Copeland
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  9. Raymond C. Sowder
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  10. Shu-jing Wei
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By acceptance of this article, the publisher or recipient acknowledges the right of the U.S. Government to retain a nonexclusive, royalty-free license in and to any copyright covering the article

Research being carried out at the Frederick Cancer Research Facility, Frederick, Maryland

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Kung, Hf., Calvert, I., Bekesi, E. et al. Phosphorylation of human interleukin-2 (IL-2). Mol Cell Biochem 89, 29–35 (1989). https://doi.org/10.1007/BF00228277

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  • DOI: https://doi.org/10.1007/BF00228277

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