Skip to main content

Advertisement

SpringerLink
Log in

Characterization of the chaperonin GroEL in Mycoplasma gallisepticum

  • Orginal Paper
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Mycoplasma gallisepticum (MG) is a common and widespread cause of chronic respiratory disease in poultry. In this study, antigenic proteins were identified from MG membrane using two-dimensional gel electrophoresis (2-DE) analysis followed by Western blot and matrix-assisted desorption/ionization time-of-flight mass spectrometry (MALDI-TOF–MS), including translation elongation factor Tu, dihydrolipoamide acetyltransferase (E2) component of pyruvate dehydrogenase complex, trigger factor, chaperone protein DnaK, heat shock protein GroEL and so on. Furthermore, recombinant MG GroEL protein was successfully expressed in E. coli BL21 (DE3) with pET-28a (+) vector and found to possess ATPase activity and contributed to the refolding of recombinant MG PrpC protein. Complement-dependent bactericidal assay indicated that the rabbit antisera against MG rGroEL had satisfactory bactericidal effect, which is similar to the chicken antisera induced by MG-inactivated vaccine, suggesting MG GroEL is a protective antigen, could be used as a novel vaccine candidate. This study is the first report of the biological characterization of chaperone GroEL protein in MG.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

MG:

Mycoplasma gallisepticum

2-DE:

Two-dimensional gel electrophoresis

MALDI-TOF–MS:

Matrix-assisted desorption/ionization time-of-flight mass spectrometry

CRD:

Chronic respiratory disease

HSPs:

Heat shock family of proteins

PMF:

Peptide mass fingerprints

MW:

Molecular weight

Pi:

Inorganic phosphate

IPTG:

Isopropyl-β-D-thiogalactopyranoside

References

  • Amemiya K, Meyers JL, Deshazer D, Riggins RN, Halasohoris S, England M, Ribot W, Norris SL, Wagg DM (2007) Detection of the host immune response to Burkholderia mallei heat-shock proteins GroEL and DnaK in a glanders patient and infected mice. Diagn Microbiol Infect Dis 59:137–147

    Article  CAS  PubMed  Google Scholar 

  • Ayalew S, Confer AW, Shrestha B, Payton ME (2012) A rapid microtiter plate serum bactericidal assay method for determining serum complement-mediated killing of Mannheimia haemolytica. J Microbiol Methods 89:99–101

    Article  CAS  PubMed  Google Scholar 

  • Bansal A, Paliwal PK, Sagi SS, Sairam M (2010) Effect of adjuvants on immune response and protective immunity elicited by recombinant Hsp60 (GroEL) of Salmonella typhi against S. typhi infection. Mol Cell Biochem 337:213–221

    Article  CAS  PubMed  Google Scholar 

  • Boguslavsky S, Menaker D, Lysnyansky I, Liu T, Levisohn S, Rosengarten R, García M, Yogev D (2000) Molecular characterization of the Mycoplasma gallisepticum pvpA gene which encodes a putative variable cytadhesin protein. Infect Immun 68:3956–3964

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Chapman E, Farr GW, Usaite R, Furtak K, Fenton WA, Chaudhuri TK, Hondorp ER, Matthews RG, Wolf SG, Yates JR, Pypaert M, Horwich AL (2006) Global aggregation of newly translated proteins in an Escherichia coli strain deficient of the chaperonin GroEL. Proc Natl Acad Sci USA 103:15800–15805

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Cleavinger CM, Kim MF, Wise KS (1994) Processing and surface presentation of the Mycoplasma hyorhinis variant lipoprotein VlpC. J Bacteriol 176:2463–2467

    PubMed Central  CAS  PubMed  Google Scholar 

  • Demina IA, Serebryakova MV, Ladygina VG, Rogova MA, Kondratov IG, Renteeva AN, Govorun VM (2010) Proteomic characterization of Mycoplasma gallisepticum nanoforming. Biochemistry 75:1252–1257

    CAS  PubMed  Google Scholar 

  • Ensgraber M, Loos M (1992) A 66-kilodalton heat shock protein of Salmonella typhimurium is responsible for binding of the bacterium to intestinal mucus. Infect Immun 60:3072–3078

    PubMed Central  CAS  PubMed  Google Scholar 

  • Fayet O, Ziegelhoffer T, Georgopoulos C (1989) The groES and groEL heat shock gene products of Escherichia coli are essential for bacterial growth at all temperatures. J Bacteriol 171:1379–1385

    PubMed Central  CAS  PubMed  Google Scholar 

  • Fujimoto A, Kosaka N, Hasegawa H, Suzuki H, Sugano S, Chiba J (2012) Enhancement of antibody responses to native G protein-coupled receptors using E. coli GroEL as a molecular adjuvant in DNA immunization. J Immunol Methods 375:243–251

    Article  CAS  PubMed  Google Scholar 

  • Goulhen F, Hafezi A, Uitto VJ, Hinode D, Nakamura R, Grenier D, Mayrand D (1998) Subcellular localization and cytotoxic activity of the GroEL-like protein isolated from Actinobacillus actinomycetemcomitans. Infect Immun 66:5307–5313

    PubMed Central  CAS  PubMed  Google Scholar 

  • Gray TE, Fersht AR (1991) Cooperativity in ATP hydrolysis by GroEL is increased by GroES. FEBS Lett 292:254–258

    Article  CAS  PubMed  Google Scholar 

  • Han X, Hu Q, Ding S, Chen W, Ding C, He L, Wang X, Ding J, Yu S (2012) Identification and immunological characteristics of chaperonin GroEL in Riemerella anatipestifer. Appl Microbiol Biotechnol 93:1197–1205

    Article  CAS  PubMed  Google Scholar 

  • Hartley MG, Green M, Choules G, Rogers D, Rees DG, Newstead S, Sjostedt A, Titball RW (2004) Protection afforded by heat shock protein 60 from Francisella tularensis is due to copurified lipopolysaccharide. Infect Immun 72:4109–4113

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hennigan SL, Driskell JD, Ferguson-Noel N, Dluhy RA, Zhao Y, Tripp RA, Krause DC (2012) Detection and differentiation of avian mycoplasmas by surface-enhanced Raman spectroscopy based on a silver nanorod array. Appl Environ Microbiol 78:1930–1935

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hu Y, Henderson B, Lund PA, Tormay P, Ahmed MT, Gurcha SS, Besra GS, Coates AR (2008) A Mycobacterium tuberculosis mutant lacking the groEL homologue cpn60.1 is viable but fails to induce an inflammatory response in animal models of infection. Infect Immun 76:1535–1546

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Huang YY, Deng JY, Gu J, Zhang ZP, Maxwell A, Bi LJ, Chen YY, Zhou YF, Yu ZN, Zhang XE (2006) The key DNA-binding residues in the C-terminal domain of Mycobacterium tuberculosis DNA gyrase A subunit (GyrA). Nucleic Acids Res 34:5650–5659

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Illingworth M, Ramsey A, Zheng Z, Chen L (2011) Stimulating the substrate folding activity of a single ring GroEL variant by modulating the cochaperonin GroES. J Biol Chem 286:30401–30408

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Khan MN, Shukla D, Bansal A, Mustoori S, Ilavazhagan G (2009) Immunogenicity and protective efficacy of GroEL (hsp60) of Streptococcus pneumoniae against lethal infection in mice. FEMS Immunol Med Microbiol 56:56–62

    Article  CAS  PubMed  Google Scholar 

  • Kobayashi K, Ehrlich SD, Albertini A, Amati G, Andersen KK, Arnaud M, Asai K, Ashikaga S, Aymerich S, O N (2003) Essential Bacillus subtilis genes. Proc Natl Acad Sci USA 100:4678–4683

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Leclerq S, Harms JS, Rosinha GM, Azevedo V, Oliveira SC (2002) Induction of a th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene. J Med Microbiol 51:20–26

    CAS  PubMed  Google Scholar 

  • Lewthwaite JC, Clarkin CE, Coates AR, Poole S, Lawrence RA, Wheeler-Jones CP, Pitsillides AA, Singh M, Henderson B (2007) Highly homologous Mycobacterium tuberculosis chaperonin 60 proteins with differential CD14 dependencies stimulate cytokine production by human monocytes through cooperative activation of p38 and ERK1/2 mitogen-activated protein kinases. Int Immunopharmacol 7:230–240

    Article  CAS  PubMed  Google Scholar 

  • Martin D, McCallum L, Glennie A, Ruijne N, Blatchford P, O’Hallahan J, Oster P (2005) Validation of the serum bactericidal assay for measurement of functional antibodies against group B meningococci associated with vaccine trials. Vaccine 23:2218–2221

    Article  CAS  PubMed  Google Scholar 

  • Mendoza JA, Warren T, Dulin P (1996) The ATPase activity of chaperonin GroEL is highly stimulated at elevated temperatures. Biochem Biophys Res Commun 229:271–274

    Article  CAS  PubMed  Google Scholar 

  • Obuchowski M, Madec E, Delattre D, Boël G, Iwanicki A, Foulger D, Seror SJ (2000) Characterization of PrpC from Bacillus subtilis, a member of the PPM phosphatase family. J Bacteriol 182:5634–5638

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Panicker IS, Kanci A, Chiu CJ, Veith PD, Glew MD, Browning GF, Markham PF (2012) A novel transposon construct expressing PhoA with potential for studying protein expression and translocation in Mycoplasma gallisepticum. BMC Microbiol 12:138

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Sielaff B, Lee KS, Tsai FT (2011) Structural and functional conservation of Mycobacterium tuberculosis GroEL paralogs suggests that GroEL1 Is a chaperonin. J Mol Biol 405:831–839

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Sinha K, Bhatnagar R (2010) GroEL provides protection against Bacillus anthracis infection in BALB/c mice. Mol Immunol 48:264–271

    Article  CAS  PubMed  Google Scholar 

  • Woo PC, Leung PK, Wong SS, Ho PL, Yuen KY (2001) groEL encodes a highly antigenic protein in Burkholderia pseudomallei. Clin Diagn Lab Immunol 8:832–836

    PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was sponsored by the Special Fund for Agro-scientific Research in the Public Interest (Grant Nos. 201303044 and 201303038). We would like to thank Prof. Howard Barry Gelberg for reading of this manuscript and professional language editing.

Conflict of interest

The authors have not declared any conflict of interest.

Author information

Authors and Affiliations

  1. Department of Avian Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No.518, Ziyue Road, Minhang District, Shanghai, 200241, People’s Republic of China

    Lei Tan, Meirong Hu, Shengqing Yu, Xin Wang, Feng Lu, Fang Liu, Xusheng Qiu, Cuiping Song, Yingjie Sun & Chan Ding

Authors
  1. Lei Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Meirong Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shengqing Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Feng Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Fang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xusheng Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Cuiping Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yingjie Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Chan Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chan Ding.

Additional information

Communicated by Erko Stackebrandt.

Lei Tan and Meirong Hu have contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tan, L., Hu, M., Yu, S. et al. Characterization of the chaperonin GroEL in Mycoplasma gallisepticum . Arch Microbiol 197, 235–244 (2015). https://doi.org/10.1007/s00203-014-1047-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00203-014-1047-2

Keywords

Search

Navigation