Isolation and Purification of an Antibacterial Protein from Immune Induced Haemolymph of American Cockroach, Periplaneta americana
-
Hamid Reza Basseri
,
Amir Dadi-Khoeni
,
Ronak Bakhtiari
,
Mandan Abolhassani
,
Reza Hajihosseini-Baghdadabadi
Abstract
Background: Antimicrobial peptides play a role as effectors substances in the immunity of vertebrate and invertebrate hosts. In the current study, antimicrobial peptide was isolated from the haemolymph of the American cockroach, Periplaneta americana.
Methods: Micrococcus luteus as Gram-positive bacteria and Escherichia coli as Gram-negative bacteria were candidate for injection. Induction was done by injecting both bacteria into the abdominal cavity of two groups of cockroaches separately. The haemolymphs were collected 24 hours after post injection and initially tested against both bacteria. Subsequently, the immune induced haemolymph was purified by high performance liquid chromatography (HPLC) to separate the proteins responsible for the antibacterial activity.
Results: The non-induced haemolymph did not show any activity against both bacteria whereas induced haemolymph exhibited high activity against M. luteus but did less against E. coli. Two fractions showed antibacterial activity against M. luteus. Finally the molecular weight of the isolated antibacterial proteins were determined as 72 kDa and 62 kDa using SDS-PAGE.
Conclusion: Induced haemolymph of American cockroaches has the ability to produce peptides to combat against Gram-positive bacteria when an immune challenge is mounted. Further work has to be done to sequence of the protein, which it would be advantageous.
Bang K, Park S, Yoo JY, Cho S (2012) Characterization and expression of at- tacin, an antibacterial protein-encoding gene, from the beet armyworm, Spodop- tera exigua (Hubner) (Insecta: Lepi- doptera: Noctuidae). Mol Biol Rep. 39:5151–5159.
Bosco-Drayon V, Poidevin M, Boneca IG, Narbonne-Reveau K, Royet J, Char- roux B ( 2012) Peptidoglycan sensing by the receptor PGRP-LE in the Dro- sophila gut induces immune responses to infectious bacteria and tolerance to microbiota. Cell Host Microbe. 12:153–165.
Brivio M, F. Moro M, Mastore M (2006) Down-regulation of antibacterial pep- tide synthesis in an insect model in- duced by the body-surface of an en- tomoparasite (Steinernema feltiae). Dev Comp Immunol. 30: 627–638.
Cociancich S, Bulet P, Hetru C, Hoffmann JA (1994) The inducible antibacterial peptides of insects. Parasitol Today.10: 132–139.
Dai H, Rayaprolu S, Gong Y, Huang R, Prakash O, Jiang H (2008) Solution structure, antibacterial activity, and ex- pression profile of Manduca sexta moricin. J Pept Sci 14: 855–863.
Eleftherianos I, Marokhazi J, Millichap PJ, Hodgkinson AJ, Sriboonlert A, ffrench- Constant RH, Reynolds SE (2006) Prior infection of Manduca sexta with non-pathogenic Escherichia coli elicits immunity to pathogenic Photorhabdus luminescens: roles of immune-related proteins shown by RNA interference. Insect Biochem Mol Biol. 36: 517–525.
Eleftherianos I, Gokcen F, Felfoldi G, Millichap PJ, Trenczek TE, ffrench- Constant RH, Reynolds SE (2007) The immunoglobulin family protein Hemo- lin mediates cellular immune respons- es to bacteria in the insect Manduca sexta. Cell Microbiol. 9: 1137–1147.
Engstrom Y (1999) Induction and regulation of antimicrobial peptides in Drosoph- ila. Dev Comp Immunol. 23: 345–358.
Faulhaber LM, Karp RD (1992) A diphasic immune response against bacteria in the American cockroach. Immunology.75: 378–381.
Fiolka MJ (2008) Immunosuppressive effect of cyclosporin A on insect humoral immune response. J Invertebr Pathol.98: 287–292.
Gao B, Zhu S (2012) Alteration of the mode of antibacterial action of a defensin by the amino-terminal loop substitution. Biochem Biophys Res Commun. 426:630–635.
Gao B, Zhu S (2013) An insect defensin- derived beta-hairpin peptide with en- hanced antibacterial activity. ACS Chem Biol. Epub ahead of print.
George JF, Karp RD, Rellahan BL, Lessard JL (1987) Alteration of the protein com- position in the haemolymph of Amer- ican cockroaches immunized with sol- uble proteins. Immunol. 62: 505–509.
Gillespie JP, Kanost MR (1997) Biology me- diators of insect immunity. Annual Re- view of Entomology. 42: 611–643.
Ha Lee J, Hee Lee I, Noda H, Mita K, Taniai K (2007) Verification of elicitor effi- cacy of lipopolysaccharides and pepti- doglycans on antibacterial peptide gene expression in Bombyx mori. Insect Bi- ochem Mol Biol. 37: 1338–1347.
Hashimoto Y, Tabuchi Y, Sakurai K, Kutsuna M, Kurokawa K, Awasaki T, Sekimizu K, Nakanishi Y, Shiratsuchi A (2009) Identification of lipoteichoic acid as a ligand for draper in the phag- ocytosis of Staphylococcus aureus by Drosophila hemocytes. J Immunol. 183:
–7460.
Hoffmann JA, Reichhart JM, Hetru C (1996) Innate immunity in higher insects. Curr Opin Immunol. 8: 8–13.
Huang Y, Lou H, Wu X, Chen Y (2008) Characterization of the BPI-like gene from a subtracted cDNA library of large yellow croaker (Pseudosciaena crocea) and induced expression by formalin- inactivated Vibrio alginolyticus and Nocardia seriolae vaccine challenges. Fish Shellfish Immunol. 25: 740–750.
Imler JL, Bulet P (2005) Antimicrobial peptides in Drosophila: structures, ac- tivities and gene regulation. Chem Immunol Allergy. 86: 1–21.
Jarosz J (1996) Anti-infective defence strate- gies and methods of escape from en- tomologic pathogens under immuno- logic control of insects. Wiad Para- zytol. 42: 3–27.
Jomori T, Natori S (1992) Function of the lipopolysaccharide-binding protein of Periplaneta americana as an opsonin. FEBS Lett. 296: 283–286.
Karp RD (1985) Preliminary characteriza- tion of the inducible humoral factor in the American cockroach (Periplaneta americana). Dev Comp Immunol. 9:569–575.
Kim CH, Shin YP, Noh MY, Jo YH, Han YS, Seong YS, Lee IH (2010) An insect multiligand recognition protein functions as an opsonin for the phag- ocytosis of microorganisms. J Biol Chem. 285: 25243–25250.
Laemmli UK (1970) Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature. 227: 680–685.
Lamberty M, Zachary D, Lanot R, Borde-reau C, Robert A, Hoffmann JA, Bulet P (2001) Insect immunity. Constitutive expression of a cysteine-rich antifun- gal and a linear antibacterial peptide in a termite insect. J Biol Chem. 276:4085–4092.
Lazzaro BP (2008) Natural selection on the Drosophila antimicrobial immune sys- tem. Curr Opin Microbiol. 11: 284–289.
Leclerc V, Reichhart JM (2004) The im- mune response of Drosophila melano- gaster. Immunol Rev. 198: 59–71.
Levy F, Rabel D, Charlet M, Bulet P, Hoffmann JA, Ehret-Sabatier L (2004) Peptidomic and proteomic analyses of the systemic immune response of Drosophila. Biochimie. 86: 607–616.
Mak P, Zdybicka-Barabas A, Cytrynska M (2010) A different repertoire of Gal- leria mellonella antimicrobial peptides in larvae challenged with bacteria and fungi. Dev Comp Immunol. 34: 1129–1136.
Rahnamaeian M, Vilcinskas A (2012) De- fense gene expression is potentiated in transgenic barley expressing antifungal peptide Metchnikowin throughout pow- dery mildew challenge. J Plant Res.125: 115–124.
Rahnamaeian M, Langen G, Imani J, Khalifa W, Altincicek B, von Wettstein D, Kogel KH, Vilcinskas A (2009) Insect peptide metchnikowin confers on bar- ley a selective capacity for resistance to fungal ascomycetes pathogens. J Exp Bot. 60: 4105–4114.
Royet J, Dziarski R (2007) Peptidoglycan recognition proteins: pleiotropic sen- sors and effectors of antimicrobial de- fences. Nat Rev Microbiol. 5: 264–277.
Royet J, Gupta D, Dziarski R (2011) Pep- tidoglycan recognition proteins: mod- ulators of the microbiome and inflam- mation. Nat Rev Immunol. 11: 837–851.
Sawa T, Kurahashi K (1999) Antimicrobial peptides/proteins-application to the ther- apy of sepsis. Masui. 48: 1186–1193.
Shen X, Ye G, Cheng X, Yu C, Altosaar I, Hu C (2010) Characterization of an abaecin-like antimicrobial peptide iden- tified from a Pteromalus puparum cDNA clone. J Invertebr Pathol. 105: 24–29.
Toke O (2005) Antimicrobial peptides: new candidates in the fight against bacterial infections. Biopolymers. 80: 717–735.
Vilcinskas A (2013) Evolutionary plasticity of insect immunity. J Insect Physiol.59: 123–129.
Wade JD, Lin F, Condie BA, Hanrieder J, Hoffmann R (2005) Designer antibac- terial peptides kill fluoroquinolone- resistant clinical isolates. J Med Chem.48: 5349–5359.
Wang Y, Jin X, Zhu J, Zeng A, Chu F, Yang X, Ma Y (2009) Expression pattern of antibacterial genes in the Musca domes- tica. Sci China C Life Sci. 52: 823–830.
Wilson R, Chen C, Ratcliffe NA (1999) Innate immunity in insects: the role of multiple, endogenous serum lectins in the recognition of foreign invaders in the cockroach, Blaberus discoidalis. J Immunol. 162: 1590–1596.
Yakovlev AY (2011) Induction of antimicro- bial peptide synthesis by the fat body cells of maggots of Calliphora vicina R.-D. (Diptera: Calliphoridae). Zh Evol Biokhim Fiziol. 47(6): 461–468.
Ye JS, Zheng XJ, Leung KW, Chen HM, Sheu FS (2004) Induction of transient ion channel-like pores in a cancer cell by antibiotic peptide. J Biochem. 136:255–259.
Yu Y, Park JW, Kwon HM, Hwang HO, Jang IH, Masuda A, Kurokawa K, Nakayama H, Lee WJ, Dohmae N, Zhang J, Lee BL(2010) Diversity of innate immune recognition mechanism for bacterial polymeric meso-diaminopimelic acid- type peptidoglycan in insects. J Biol Chem. 285: 32937–32945.
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| Issue | Vol 10 No 4 (2016) | |
| Section | Original Article | |
| Keywords | ||
| American Cockroach Antibacterial Protein Isolation Micrococcus luteus Escherichia coli | ||
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