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A Low-Molecular-Weight Compound Derived from Human Leukocytes Determines a Bactericidal Activity of the Interferon Preparation

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Abstract

The aim of this study was to characterize the structure and mode of action of antimicrobials derived from a commercial preparation of alfa-interferon. By combination of semi-preparative/analytical reversed-phase high-performance liquid chromatography, we isolated and purified a novel active substance based on carbohydrate with a complex of amino acids, which determines antimicrobial property of commercial preparation of interferon. A size-exclusion chromatography was performed and LC/ESI-MS revealed molecular masses of active substance were in the range of 180–249 Da. Edman sequencing identified phenylthiohydantoin (PTH) derivatives which consisted a set of preliminary (Asp, Glu, Gly, and Ala) and minor amino acids (Leu and Thr) at equimolar ratio. Thus, the purified active substance is a compound containing the complex of amino acids connected with carbohydrate background and called leucidin. Leucidin demonstrated antimicrobial activity against the model Escherichia coli (E. coli) K12 strain at a minimal inhibitory concentration of 20 μg mL−1. The revealed antimicrobial mechanism of action is associated with violation of the bacterial cell wall leading to a SOS response and bacterial autolysis. Despite the preliminary nature of the results, obtained data allowed us to discover the previously unknown leukocyte-derived antimicrobial molecules.

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Acknowledgments

The authors are grateful to Dr. Nikolay Yu. Tretyakov (Institute of Chemistry, Tyumen State University); we are grateful to Dr. Ilya V. Manukhov (Moscow Institute of Physics and Technology (State University)) for providing bacterial reporter strains; we are grateful to Dr. Alexander A. Kolobov (Research Institute of Highly Pure Biopreparations, St. Peterburg, Russian Federation) for providing indolicidin preparation. We are grateful to Nadezhda N. Zhuravleva (Center for Academic Writing, Tyumen State University) for language assistance.

The research was partially performed using the equipment of the Research Resource Center “Natural Resource Management and Physico-Chemical Research” of Tyumen State University; and Federal Research Centre of Biological Systems and Agro-technologies, RAS.

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Authors and Affiliations

  1. Tyumen State University, Volodarsky st 6, Tyumen, Russian Federation

    A. S. Vasilchenko

  2. Institute of Cellular and Intracellular Symbiosis, RAS, Pionerskaya st 11, Orenburg, Russian Federation

    V. A. Gritsenko

  3. Federal Research Centre of Biological Systems and Agro-technologies, RAS, Yanvarya st 9, Orenburg, Russia

    D. B. Kosyan

  4. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, RAS, ul. Miklukho-Maklaya, 16/10, Moscow, Russian Federation

    E. A. Rogozhin

  5. Gause Institute of New Antibiotics, ul. Bolshaya Pirogovskaya, 11, Moscow, Russian Federation

    E. A. Rogozhin

Authors
  1. A. S. Vasilchenko
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  2. V. A. Gritsenko
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  3. D. B. Kosyan
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  4. E. A. Rogozhin
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Correspondence to A. S. Vasilchenko.

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Vasilchenko, A.S., Gritsenko, V.A., Kosyan, D.B. et al. A Low-Molecular-Weight Compound Derived from Human Leukocytes Determines a Bactericidal Activity of the Interferon Preparation. Probiotics & Antimicro. Prot. 11, 999–1008 (2019). https://doi.org/10.1007/s12602-018-9463-2

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