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The LKEKK synthetic peptide as a ligand of rat intestinal epithelial cell membranes

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Abstract

A tritium-labeled synthetic LKEKK pentapeptide corresponding to the sequences 16–20 of human thymosin-α1 and 131–135 of human interferon-α2 was obtained with a specific activity of 28 Ci/mmol. [3H]LKEKK was found to bind with high affinity (K d 3.7 ± 0.3 nM) to the membranes isolated from epithelial cells of rat small intestinal mucosa. The trypsin treatment of the membranes did not affect the binding, thus supporting the nonprotein nature of the peptide receptor. The binding of the labeled peptide was inhibited by unlabeled thymosin-α1, interferon-α2, and cholera toxin B subunit (K i 4.2 ± 0.4, 3.5 ± 0.3, and 4.7 ± 0.3 nM respectively). The pentapeptide did not affect the adenylate cyclase activity within the concentration range of 1–1000 nM.

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Abbreviations

cAMP:

adenosine 3′,5′-cyclophosphate

CT-A and CT-B:

cholera toxin A and B subunits

IL:

interleukin

IFN:

interferon

TM-α1 :

thymosin α1

TNF:

tumor necrosis factor

MyD88:

the protein of the myeloid differentiation primary response 88

TIR domain:

Toll/IL-1 receptor resistant domain

TCA:

trichloroacetic acid

TLRs:

Toll-like receptors

TRIF:

the adaptor protein containing the TIR domain

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Authors and Affiliations

  1. Branch of Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, pr. Nauki 6, Pushchino, Moscow oblast, 142290, Russia

    E. V. Navolotskaya, V. B. Sadovnikov, D. V. Zinchenko & V. I. Vladimirov

  2. Institute of Molecular Genetics of the Russian Academy of Science, pl. akad. Kurchatova 2, Moscow, 123182, Russia

    Y. A. Zolotarev

  3. State Research Center for Institute of Highly Pure Biopreparations of Federal Biomedical Agency, ul. Pudozhskaya 7, St. Petersburg, 197110, Russia

    A. A. Kolobov

Authors
  1. E. V. Navolotskaya
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  2. V. B. Sadovnikov
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  3. D. V. Zinchenko
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  4. V. I. Vladimirov
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  5. Y. A. Zolotarev
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  6. A. A. Kolobov
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Correspondence to E. V. Navolotskaya.

Additional information

Original Russian Text © E.V. Navolotskaya, V.B. Sadovnikov, D.V. Zinchenko, V.I. Vladimirov, Y.A. Zolotarev, A.A. Kolobov, 2016, published in Bioorganicheskaya Khimiya, 2016, Vol. 42, No. 5, pp. 533–538.

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Navolotskaya, E.V., Sadovnikov, V.B., Zinchenko, D.V. et al. The LKEKK synthetic peptide as a ligand of rat intestinal epithelial cell membranes. Russ J Bioorg Chem 42, 479–483 (2016). https://doi.org/10.1134/S1068162016050137

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  • DOI: https://doi.org/10.1134/S1068162016050137

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