Email this article FUNCTIONAL EFFECTS OF θ-DEFENSINS FROM BLOOD LEUKOCYTES OF BABOON PAPIO HAMADRYAS
- Authors: Tsvetkova E.V.1, Aleshina G.M.1, Leonovа L.E.2, Shamova O.V.1, Romanovskaya E.V.2, Kokryakov V.N.1
-
Affiliations:
- Saint-Petersburg State University, Research Institute of Experimental Medicine
- Saint-Petersburg State University
- Issue: Vol 95, No 5 (2018)
- Pages: 66-73
- Section: ORIGINAL RESEARCHES
- URL: https://microbiol.crie.ru/jour/article/view/310
- DOI: https://doi.org/10.36233/0372-9311-2018-5-66-73
- ID: 310
Cite item
Full Text
Abstract
Aim. Study the functional properties of cationic antimicrobial peptides 0-defensins isolated from baboon Papio hamadryas blood leukocytes. Materials and methods. Baboon 0-defensins were extracted from leukocyte mass using ultrafiltration, preparative electrophoresis and reverse phase high performance liquid chromatography. The test microorganisms used were Gram-negative bacteria Escherichia coli, Gram-positive bacteria Listeria monocytogenes and Staphylococcus aureus, and fungi Candida albicans. Minimal inhibitory concentrations (MICs) were determined by testing serial dilutions of the test peptides by radial diffusion in agarose gel. Microbicidal action was evaluated by counting surviving colony forming units after incubation microorganisms with the peptides. The 0-defensins influence on E.coli membrane permeability was assessed using chromogenic markers o-nirofenil- β-D-galactopyranoside and nitrocefin. Results. The analysis of the 0-defensins antimicrobial properties showed that they produce antimicrobial activity against test microorganisms, exhibiting bactericidal and fungicidal effects at micromolar concentrations. We studied the influence of different environmental conditions (low and high ionic strength, blood serum in medium) for antimicrobial activity. It is shown that 0-defensins have the ability to increase the outer membran of E.coli permeability, however, in contrast to 0-defensins have no noticeable influence on the inner membrane permeability. Conclusion. Baboon 0-defensins isolated from blood leukocytes are effective antimicrobial agents with a broad spectrum of microbicidal action. 0-Defensins baboon unlike 0-defensins exhibit antimicrobial activity in environments with high ionic strength. 0-Defensins increase the outer membran of E.coli permeability but not the cytoplasmic membrane, suggesting that the mechanism of antimicrobial effect of these peptides other than 0-defensins.
About the authors
E. V. Tsvetkova
Saint-Petersburg State University, Research Institute of Experimental Medicine
Author for correspondence.
Email: noemail@neicon.ru
Russian Federation
G. M. Aleshina
Saint-Petersburg State University, Research Institute of Experimental Medicine
Email: noemail@neicon.ru
Russian Federation
L. E. Leonovа
Saint-Petersburg State University
Email: noemail@neicon.ru
Russian Federation
O. V. Shamova
Saint-Petersburg State University, Research Institute of Experimental Medicine
Email: noemail@neicon.ru
Russian Federation
E. V. Romanovskaya
Saint-Petersburg State University
Email: noemail@neicon.ru
Russian Federation
V. N. Kokryakov
Saint-Petersburg State University, Research Institute of Experimental Medicine
Email: noemail@neicon.ru
Russian Federation
References
- Abuja P.M., Zenz A., Trabi M. et al. The cyclic antimicrobial peptide RTD-1 induces stabilized lipid-peptide domains more efficiently than its open-chain analogue. FEBS Letters. 2004, 566: 301-306.
- Buffy J.J., McCormick M.J., Wi S. et al. Solid-state NMR investigation of selective perturbation of lipid bilayers by cyclic antimicrobial peptide RTD-1. Biochemistry. 2004, 43: 9800-9812.
- Garcia A.E., Usapay G., Tran PA. et al. Isolation, synthesis, and antimicrobial activities of naturally occurring 0-defensin isoforms from baboon leukocytes. Infect. Immun. 2008, 76: 5883-5891.
- Lehrer R., Barton A., Daher K. et al. Interaction of human defensins with Escherichia coli. Mechanism of bactericidal activity. J. Clin. Invest. 1989, 84: 553-561.
- Lehrer R.I., Rosenman M., Harwig S.S.L. et al. Ultrasensitive assays for endogenous antimicrobial polypeptides. J. Immunol. Methods. 1991, 137:167-173.
- Lehrer R.I., Lu W. а-Defensins in human innate immunity. Immunol. Rev. 2012, 245: 84-112.
- Pace C.N., Vajdos F., Fee L. et al. How to measure and predict the molar absorption coefficient of a protein. Protein Sci. 1995, 4: 2411-2423.
- Stegemann C., Tsvetkova E.V., Aleshina G.M. et al. De novo sequencing of two new cyclic theta-defensins from baboon (Papio hamadryas) leukocytes by matrix-assisted laser desorption/ ionization mass spectrometry. Rapid Commun. Mass Spectrom. 2010, 24:599-604.
- Tang Y.Q., Yuan J., Osapay G. et al. A cyclic antimicrobial peptide produced in primate leukocytes by the ligation of two truncated alpha-defensins. Science. 1999, 286: 498-502.
- Tran D., Tran P., Roberts K. et al. Microbicidal properties and cytocidal selectivity of rhesus macaque theta defensins. Antimicrob. Agents Chemother. 2008, 52: 944-953.
- Tsvetkova E.V., Aleshina G.M., Shamova O.V. et al. а-Defensins from blood leukocytes of the monkey Papio hamadryas. Biochemistry (Moskow). 2006, 71: 879-883.
- Tsvetkova E.V., Leonova L.E., Aleshina G.M. et al. Antimicrobial effects of а-defensins from leukocytes of the hamadryas baboon Papio hamadryas. J. Evolutionary Biochemistry Physiology. 2016, 52(2): 133-140.
- Varkey J., Nagaraj R. Antibacterial activity of human neutrophil defensin HNP-1 analogs without cysteines. Antimicrob. Agents Chemother. 2005, 49: 4561-4566.
- Weiss T.M., Yang L., Ding L. et al. Two states of cyclic antimicrobial peptide RTD-1 in lipid bilayers. Biochemistry. 2002, 41: 10070-10076.
- Yamaguchi S., Hong T., Waring A. et al. Solid-state NMR investigations of peptide-lipid interaction and orientation of a в-sheet antimicrobial peptide, protegrin. Biochemistry. 2002, 41: 9852-9862.
Supplementary files
