Ukr.Biochem.J. 2019; Volume 91, Issue 1, Jan-Feb, pp. 65-73
doi: https://doi.org/10.15407/ubj91.01.065
HBD-2 interactions with erythrocyte membranes in vitro
M. V. Makarenko1, D. O. Semeniuk2, I. O. Starenka2, A. P. Pogribna3,
I. V. Sokol4, L. I. Martinova1, D. O. Govsieiev4
1Institute of Postgraduate Education,
Bogomolets National Medical University, Kyiv Ukraine;
2ESC Institute of Biology and Medicine,
Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: dmyt.semenyuk@gmail.com;
3Institute of Molecular Biology and Genetics, NAS of Ukraine, Kyiv;
4Kyiv City Maternity Hospital No 5
Received: 24 September 2018; Accepted: 13 December 2018
HBD-2 – a member of β-defensin family of antimicrobial peptides – is known to permeabilize cell membranes of susceptible cells, but the mechanism of such interactions is poorly understood. In our study, we have used a hemolytic model to explore the kinetic properties of HBD-2 interactions with membranes of human erythrocytes. We ran hemolytic assays with a wide range of both HBD-2 and erythrocyte concentrations, as well as varying pH values, incubation times, and osmotic strengths; each in the presence or the absence of inhibitory substances such as proteins and salts. The results show that HBD-2 cell membrane permeabilization is both dose- and time-dependent (with plateau effect observed in each case), and inversely dependent on erythrocyte concentration. HBD-2 interactions with cell membranes highly depend on pH value and the presence of inhibitors but are not affected by tested osmotic strength range. Our findings suggest that interactions of HBD-2 with cell membranes are mainly electrostatic in nature and are limited by released cell content. We have developed a speculative model of such interaction based on our results.
Keywords: hemolytic activity, human beta-defensin 2, membrane permeabilization
References:
- Bals R, Wang X, Wu Z, Freeman T, Bafna V, Zasloff M, Wilson JM. Human beta-defensin 2 is a salt-sensitive peptide antibiotic expressed in human lung. J Clin Invest. 1998 Sep 1;102(5):874-80. PubMed, PubMedCentral, CrossRef
- Harder J, Bartels J, Christophers E, Schröder JM. A peptide antibiotic from human skin. Nature. 1997 Jun 26;387(6636):861. PubMed, CrossRef
- Schröder JM, Harder J. Human beta-defensin-2. Int J Biochem Cell Biol. 1999 Jun;31(6):645-51. PubMed
- Sørensen OE, Thapa DR, Rosenthal A, Liu L, Roberts AA, Ganz T. Differential regulation of beta-defensin expression in human skin by microbial stimuli. J Immunol. 2005 Apr 15;174(8):4870-9. PubMed, CrossRef
- Joly S, Maze C, McCray PB Jr, Guthmiller JM. Human beta-defensins 2 and 3 demonstrate strain-selective activity against oral microorganisms. J Clin Microbiol. 2004 Mar;42(3):1024-9. PubMed, PubMedCentral, CrossRef
- Gwyer Findlay E, Currie SM, Davidson DJ. Cationic host defence peptides: potential as antiviral therapeutics. BioDrugs. 2013 Oct;27(5):479-93. PubMed, PubMedCentral, CrossRef
- Nguyen LT, Haney EF, Vogel HJ. The expanding scope of antimicrobial peptide structures and their modes of action. Trends Biotechnol. 2011 Sep;29(9):464-72. PubMed, CrossRef
- Huang LC, Redfern RL, Narayanan S, Reins RY, McDermott AM. In vitro activity of human beta-defensin 2 against Pseudomonas aeruginosa in the presence of tear fluid. Antimicrob Agents Chemother. 2007 Nov;51(11):3853-60. PubMed, PubMedCentral, CrossRef
- Wimley WC, Selsted ME, White SH. Interactions between human defensins and lipid bilayers: evidence for formation of multimeric pores. Protein Sci. 1994 Sep;3(9):1362-73. PubMed, PubMedCentral, CrossRef
- Hoover DM, Rajashankar KR, Blumenthal R, Puri A, Oppenheim JJ, Chertov O, Lubkowski J. The structure of human beta-defensin-2 shows evidence of higher order oligomerization. J Biol Chem. 2000 Oct 20;275(42):32911-8. PubMed, CrossRef
- Bauer F, Schweimer K, Klüver E, Conejo-Garcia JR, Forssmann WG, Rösch P, Adermann K, Sticht H. Structure determination of human and murine beta-defensins reveals structural conservation in the absence of significant sequence similarity. Protein Sci. 2001 Dec;10(12):2470-9. PubMed, PubMedCentral, CrossRef
- Pogribnoy PV, Lisovskiy IL, Markeeva NV, Shnitsar VM, Zinchenko II, Soldatkina MA. Production of recombinant of hbd-2-human antimicrobial peptide expressed in cervical and vulval cancer. Exp Oncol. 2003 March; 25(1):36-39.
- Greenberg CS, Craddock PR. Rapid single-step membrane protein assay. Clin Chem. 1982 Jul;28(7):1725-6. PubMed
- Harder J, Bartels J, Christophers E, Schroder JM. Isolation and characterization of human beta -defensin-3, a novel human inducible peptide antibiotic. J Biol Chem. 2001 Feb 23;276(8):5707-13. PubMed, CrossRef
- Nakayama K, Jia YX, Hirai H, Shinkawa M, Yamaya M, Sekizawa K, Sasaki H. Acid stimulation reduces bactericidal activity of surface liquid in cultured human airway epithelial cells. Am J Respir Cell Mol Biol. 2002 Jan;26(1):105-13. PubMed, CrossRef
- Tomita T, Hitomi S, Nagase T, Matsui H, Matsuse T, Kimura S, Ouchi Y. Effect of ions on antibacterial activity of human beta defensin 2. Microbiol Immunol. 2000;44(9):749-54. PubMed, CrossRef
