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Interaction of cholera toxin B-subunit with human T-lymphocytes

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Abstract

In this work, 125I-labeled cholera toxin B-subunit (CT-B) (specific activity 98 Ci/mmol) was prepared, and its high-affinity binding to human blood T-lymphocytes (K d = 3.3 nM) was determined. The binding of the 125I-labeled CT-B was inhibited by unlabeled interferon-α2 (IFN-α2), thymosin-α1 (TM-α1), and by the synthetic peptide LKEKK, which corresponds to sequences 16-20 of human TM-α1 and 131-135 of IFN-α2 (K i 0.8, 1.2, and 1.6 nM, respectively), but was not inhibited by the unlabeled synthetic peptide KKEKL with inverted sequence (K i > 1 μM). In the concentration range of 10-1000 nM, both CT-B and peptide LKEKK dose-dependently increased the activity of soluble guanylate cyclase (sGC) but did not affect the activity of membrane-bound guanylate cyclase. The KKEKL peptide tested in parallel did not affect sGC activity. Thus, the CT-B and peptide LKEKK binding to a common receptor on the surface of T-lymphocytes leads to an increase in sGC activity.

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Abbreviations

cGMP:

cyclic guanosine monophosphate

CT-B:

cholera toxin B-subunit

HPLC:

high performance liquid chromatography

IFN:

interferon

IL:

interleukin

iNOS:

inducible NO-synthase

K d :

equilibrium dissociation constant

K i :

inhibition constant

mGC:

membrane-bound guanylate cyclase

PMSF:

phenylmethylsulfonyl fluoride

sGC:

soluble guanylate cyclase

TM-α1:

thymosin-α1

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Authors and Affiliations

  1. Branch of Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 142290, Pushchino, Moscow Region, Russia

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

  2. Institute of Molecular Genetics, Russian Academy of Sciences, 123182, Moscow, Russia

    Y. A. Zolotarev

  3. Shemyakin−Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia

    V. M. Lipkin

  4. University of Turku, Turku, 20500, Finland

    V. P. Zav’yalov

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. Y. A. Zolotarev
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  5. V. M. Lipkin
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  6. V. P. Zav’yalov
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Correspondence to E. V. Navolotskaya.

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Original Russian Text © E. V. Navolotskaya, V. B. Sadovnikov, D. V. Zinchenko, Y. A. Zolotarev, V. M. Lipkin, V. P. Zav'yalov4, 2017, published in Biokhimiya, 2017, Vol. 82, No. 9, pp. 1330-1337.

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Navolotskaya, E.V., Sadovnikov, V.B., Zinchenko, D.V. et al. Interaction of cholera toxin B-subunit with human T-lymphocytes. Biochemistry Moscow 82, 1036–1041 (2017). https://doi.org/10.1134/S0006297917090061

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  • DOI: https://doi.org/10.1134/S0006297917090061

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