ABC  Vol.3 No.1 , February 2013
Amino terminus mutant OmpA from an isolated antibiotic resistant Escherichia coli still possess resistance to environmental stresses

Antibiotic resistant Escherichia coli strains are becoming more common recently. OmpA is a very important antigen protein of E. coli, which consists of two separate domains, N-terminal and C-terminal domain. The N-terminal domain contains eight β- barrel regions that plays important roles in the multifaceted functions of OmpA. In the present study, we cloned a mutant OmpA gene from a multi-antibiotic resistant E. coli strain. Sequence analysis indicated that the N-terminal DNA sequence of the mutant OmpA shared 81.05% homology with the modeled OmpA from E. coli K12 and the N-terminal amino acid sequence of the mutant OmpA was 81.22% identical to that of the E. coli K12 OmpA. Moreover, several amino acids located in the β-barrel region were mutated. The mutant OmpA was expressed in BL21 suggested by SDS-PAGE. Resistance to environmental stress assay indicated that the N-terminus mutant OmpA still possessed excellent activities in pH, temperature and osmotic pressure resistance. Our pre- sent study may supply insights into better and deeper understand the relationships between OmpA N-terminal regions and its functions in environmental stress conditions and the mechanisms on antibiotic resistance of E. coli.

Cite this paper
Zhao, Z. , Nie, X. , Li, Z. , Zhang, Z. , Ding, J. and Xie, W. (2013) Amino terminus mutant OmpA from an isolated antibiotic resistant Escherichia coli still possess resistance to environmental stresses. Advances in Biological Chemistry, 3, 108-113. doi: 10.4236/abc.2013.31014.
[1]   Nicholson, T.F., Watts, K.M. and Hunstad, D.A. (2009) OmpA of uropathogenic Escherichia coli promotes postinvasion pathogenesis of cystitis. Infection and Immunity, 12, 5245-5251. doi:10.1128/IAI.00670-09

[2]   Martinez, J.L. and Baquero, F. (2002) Interactions among strategies associated with bacterial infection: Pathogenicity, epidemicity, and antibiotic resistance. Clinical Microbiology Reviews, 4, 647-679. doi:10.1128/CMR.15.4.647-679.2002

[3]   Gohre, V., Jones, A.M., Sklenar, J., et al. (2012) Molecular crosstalk between PAMP-triggered immunity and photosynthesis. Molecular Plant-Microbe Interactions, 8, 1083-1092. doi:10.1094/MPMI-11-11-0301

[4]   Saenz, Y., Brinas, L., Dominguez, E., et al. (2004) Mechanisms of resistance in multiple-antibiotic-resistant Escherichia coli strains of human, animal, and food origins. Antimicrobial Agents and Chemotherapy, 10, 3996 4001. doi:10.1128/AAC.48.10.3996-4001.2004

[5]   Desclos-Theveniau, M., Arnaud, D., Huang, T.Y., et al. (2012) The Arabidopsis lectin receptor kinase LecRK V.5 represses stomatal immunity induced by Pseudomonas syringaepv. tomato DC3000. PLOS Pathogens, 2, e1002513. doi:10.1371/journal.ppat.1002513

[6]   Hagan, C.L., Kim, S. and Kahne, D. (2010) Reconstitution of outer membrane protein assembly from purified components. Science, 5980, 890-892. doi:10.1126/science.1188919

[7]   Sugawara, E. and Nikaido, H. (2012) OmpA is the principal nonspecific slow porin of Acinetobacter baumannii. Journal of Bacteriology, 15, 4089-4096. doi:10.1128/JB.00435-12

[8]   Smith, S.G., Mahon, V., Lambert, M.A., et al. (2007) A molecular Swiss army knife: OmpA structure, function and expression. FEMS Microbiology Letters, 1, 1-11. doi:10.1111/j.1574-6968.2007.00778.x

[9]   Yagi, K. (2007) Applications of whole-cell bacterial sensors in biotechnology and environmental science. Applied Microbiology and Biotechnology, 6, 1251-1258. doi:10.1007/s00253-006-0718-6

[10]   Kleinschmidt, J.H. (2003) Membrane protein folding on the example of outer membrane protein A of Escherichia coli. Cellular and Molecular Life Sciences, 8, 1547-1558. doi:10.1007/s00018-003-3170-0

[11]   Mohan Nair, M.K. and Venkitanarayanan, K. (2007) Role of bacterial OmpA and host cytoskeleton in the invasion of human intestinal epithelial cells by Enterobacter sa kazakii. Pediatric Research, 6, 664-669. doi:10.1203/PDR.0b013e3181587864

[12]   Barrios, A.F., Zuo, R., Ren, D., et al. (2006) Hha, YbaJ, and OmpA regulate Escherichia coli K12 biofilm formation and conjugation plasmids abolish motility. Biotechnology and Bioengineering, 1, 188-200. doi:10.1002/bit.20681

[13]   Bartra, S.S., Gong, X., Lorica, C.D., et al. (2012) The outer membrane protein A (OmpA) of Yersinia pestis promotes intracellular survival and virulence in mice. Microbial Pathogenesis, 1, 41-46. doi:10.1016/j.micpath.2011.09.009

[14]   Pore, D., Mahata, N. and Chakrabarti, M.K. (2012) Outer membrane protein A (OmpA) of Shigella flexneri 2a links innate and adaptive immunity in a TLR2-dependent manner and involvement of IL-12 and nitric oxide. Journal of Biological Chemistry, 15, 12589-12601. doi:10.1074/jbc.M111.335554

[15]   Maruvada, R. and Kim, K.S. (2012) IbeA and OmpA of Escherichia coli K1 exploit Rac1 activation for invasion of human brain microvascular endothelial cells. Infection and Immunity, 6, 2035-2041. doi:10.1128/IAI.06320-11

[16]   Pautsch, A. and Schulz, G.E. (1998) Structure of the outer membrane protein A transmembrane domain. Nature Structural & Molecular Biology, 11, 1013-1017. doi:10.1038/2983

[17]   Arora, A., Abildgaard, F., Bushweller, J.H., et al. (2001) Structure of outer membrane protein A transmembrane domain by NMR spectroscopy. Nature Structural & Molecular Biology, 4, 334-338. doi:10.1038/86214

[18]   Ried, G., Koebnik, R., Hindennach, I., et al. (1994) Membrane topology and assembly of the outer membrane protein OmpA of Escherichia coli K12. Molecular Genetics and Genomics, 2, 127-135.

[19]   Sugawara, E., Steiert, M., Rouhani, S., et al. (1996) Secondary structure of the outer membrane proteins OmpA of Escherichia coli and OprF of Pseudomonas aeruginosa. Journal of Bacteriology, 20, 6067-6069.

[20]   Tamm, L.K., Arora, A. and Kleinschmidt, J.H. (2001) Structure and assembly of beta-barrel membrane proteins. Journal of Biological Chemistry, 35, 32399-32402. doi:10.1074/jbc.R100021200

[21]   Sugawara, E. and Nikaido, H. (1994) OmpA protein of Escherichia coli outer membrane occurs in open and closed channel forms. Journal of Biological Chemistry, 27, 17981-17987.

[22]   Saint, N., El Hamel, C., De, E., et al. (2000) Ion channel formation by N-terminal domain: A common feature of OprFs of Pseudomonas and OmpA of Escherichia coli. FEMS Microbiology Letters, 2, 261-265. doi:10.1111/j.1574-6968.2000.tb09296.x

[23]   Soulas, C., Baussant, T., Aubry, J.P., et al. (2000) Outer man macrophages. Journal of Immunology, 5, 2335-2340.

[24]   Sato, M., Machida, K., Arikado, E., et al. (2000) Expres sion of outer membrane proteins in Escherichia coli growing at acid pH. Applied and Environmental Microbiology, 3, 943-947. doi:10.1128/AEM.66.3.943-947.2000

[25]   Begic, S. and Worobec, E.A. (2006) Regulation of Serra tia marcescens ompF and ompC porin genes in response to osmotic stress, salicylate, temperature and pH. Micro biology, Pt 2, 485-491. doi:10.1099/mic.0.28428-0

[26]   Hutsul, J.A. and Worobec, E. (1997) Molecular charac terization of the Serratia marcescens OmpF porin, and analysis of S. marcescens OmpF and OmpC osmoregulation. Microbiology, 143, 2797-2806. doi:10.1099/00221287-143-8-2797

[27]   Beher, M.G., Schnaitman, C.A. and Pugsley, A.P. (1980) Major heat-modifiable outer membrane protein in gram negative bacteria: Comparison with the ompA protein of Escherichia coli. Journal of Bacteriology, 2, 906-913.