Purification and parcial characterization of a hemagglutinating factor (HAF): a possible adhesive factor of the diffuse adherent of Escherichia coli (DAEC)

Tomomasa, Yano; Catani, Cleide Ferreira; Arita, Michiko; Honda, Takeshi; Miwatani, Toshio

doi:10.1590/S0036-46651996000600003

Abstracts

The mannose-resistant hemagglutinating factor (HAF) was extracted and purified from a diffuse adherent Escherichia coli (DAEC) strain belonging to the classic enteropathogenic E. coli (EPEC) serotype (0128). The molecular weight of HAF was estimated to be 18 KDa by SDS-PAGE and 66 KDa by Sephadex G100, suggesting that the native form of HAF consists of 3-4 monomeric HAF. Gold immunolabeling with specific HAF antiserum revealed that the HAF is not a rigid structure like fimbriae on the bacterial surface. The immunofluorescence test using purified HAF on HeLa cells, in addition to the fact that the HAF is distributed among serotypes of EPEC, suggests that HAF is a possible adhesive factor of DAEC strains

Escherichia coli; Diffuse adherence; Hemagglutinating factor (HAF); adhesive factor

O fator hemaglutinante (HAF) foi extraído e purificado a partir de amostra de Escherichia coli de aderência difusa (DAEC), pertencente ao sorogrupo 0128 de E. coli enteropatogênica clássica (EPEC). O peso molecular do HAF foi estimado cm 18.000 Da usando SDS-PAGE e 66.000 Da empregando Sephadex G 100, o que sugere a estrutura do HAF consistir de 3 a 4 subunidades. O emprego da técnica "gold immunolabeling" com antissoro específico anti-HAF revelou que este fator não é uma estrutura do tipo fímbria na superfície da bactéria. O resultado do teste de imunofluorescência usando HAF purificado em células HeLa, além do fato deste fator estar presente entre os sorotipos de EPEC, sugerem que o HAF seja um possível fator de aderência das amostras de DAEC.

MICROBIOLOGY

Purification and parcial characterization of a hemagglutinating factor (HAF): a possible adhesive factor of the diffuse adherent of Escherichia coli (DAEC)

Purificação e caracterização parcial de um fator hemaglutinante (HAF): uma possível adesina de Escherichia coli do tipo aderência difusa (DAEC)

Tomomasa Yano^I; Cleide Ferreira Catani^I; Michiko Arita^II; Takeshi Honda^II; Toshio Miwatani^II

^IDepartment of Microbiology and Immunology, Institute of Biology, University of Campinas, Campinas, SP, Brazil

^IIDepartment of Bacterial Infections, Research Institute for Microbiol Diseases, Osaka University, Osaka, Japan

^{Correspondence to} Correspondence to: T. Yano Caixa Postal 6109 13081-970 Campinas, SP, Brasil Tel.:(+55-19) 2397945; Fax (+55-19) 2393124

SUMMARY

The mannose-resistant hemagglutinating factor (HAF) was extracted and purified from a diffuse adherent Escherichia coli (DAEC) strain belonging to the classic enteropathogenic E. coli (EPEC) serotype (0128). The molecular weight of HAF was estimated to be 18 KDa by SDS-PAGE and 66 KDa by Sephadex G100, suggesting that the native form of HAF consists of 3-4 monomeric HAF. Gold immunolabeling with specific HAF antiserum revealed that the HAF is not a rigid structure like fimbriae on the bacterial surface. The immunofluorescence test using purified HAF on HeLa cells, in addition to the fact that the HAF is distributed among serotypes of EPEC, suggests that HAF is a possible adhesive factor of DAEC strains.

Keywords:Escherichia coli; Diffuse adherence; Hemagglutinating factor (HAF); adhesive factor.

RESUMO

O fator hemaglutinante (HAF) foi extraído e purificado a partir de amostra de Escherichia coli de aderência difusa (DAEC), pertencente ao sorogrupo 0128 de E. coli enteropatogênica clássica (EPEC). O peso molecular do HAF foi estimado cm 18.000 Da usando SDS-PAGE e 66.000 Da empregando Sephadex G 100, o que sugere a estrutura do HAF consistir de 3 a 4 subunidades. O emprego da técnica "gold immunolabeling" com antissoro específico anti-HAF revelou que este fator não é uma estrutura do tipo fímbria na superfície da bactéria. O resultado do teste de imunofluorescência usando HAF purificado em células HeLa, além do fato deste fator estar presente entre os sorotipos de EPEC, sugerem que o HAF seja um possível fator de aderência das amostras de DAEC.

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

ACKNOWLEDGEMENTS

This work was supported by grants from the Conselho Nacional de Pesquisa e Desenvolvimento (CNPq) and from the Japan Society for the Promotion of Science (JSPS).

Recebido para publicação em 23/07/1996

Aceito para publicação em 13/12/1996

1. BALDINI, M.M.; KAPER, J.B.; LEVINE, M.M. & CANDY, D.C.A. - Plasmid-mediated adhesion in enteropathogenic Escheridia coli. J. pediat. Gastroent. Nutr., 2:534-538, 1983.
2. BALDINI, M.M.; NATARO, J.P. & KAPER, J.B. - Localization of a determinant for HEp-2 adherence by enterophatogenic Escherichia coli. Infect. Immun., 52:334-336, 1986.
3. BILGE, S.S.; LAUSEN, C.R.; LAU, W. & MOSELEY, S.L. - Molecular characterization of a fimbrial adhesin, F1845, mediating diffuse adherence of diarrhea-associated Escherichia coli to HEp-2 cells. J. Bact., 171:4281-4289, 1989.
4. DONNERBERG, M.S. & KAPER, J.B. - Enteropathogenic Escherichia coli. Infect. Immun., 60:3953-3961, 1992.
5. CATANI, C.F.; YAMADA, A.T.; VIDOTTO, M.C. & YANO, T. -Adhesion of bovine enterotoxigenic Escherichia coli (ETEC) by type l-like fimbriae. FEMS Microbiol. Lett., 137:241-245, 1996.
6. EVANS, D.G.; EVANS Jr., D.J. & TJOA, W. - Haemagglutination of human group A erythrocythes by enterotoxigenic and enteropathogenic Escherichia coli isolated from adults with diarrhea: correlation with colonization factor. Infect. Immun., 18:330-337, 1977.
7. GIRON, J.A.; ASY, H.O. & SCHOOLNIK, G.K. - An inducible bundle-forming pilus of enteropathogenic Escherichia coli. Science, 254:710-713, 1991.
8. GIRON, J.A.; JONES, T.; MILLÁN-VELASCO, F. et al. - Diffuse-adhering Escherichia coli (DAEC) as a putative cause of diarrhea in Mayan children in Mexico. J. infect. Dis., 163:507-513, 1991.
9. GIRON, J.A.; ASY, H.O. & SCHOOLNIK, G.K. - Characterization of fimbriae produced by enteropathogenic Escherichia coli. J. Bact., 175:7391-7403, 1993.
10. GOMES, T.A.T.; BLAKE, P.A. & TRABULSI, L.R. - Prevalence of Escherichia coli strains with localized diffuse and aggregative adherence to HeLa cells in infants with diarrhea and matched controls. J. clin. Microbiol., 27:266-269, 1989.
11. HARTREE, E.F. -Determination of protein. Anal. Biochem., 48:422-427, 1972.
12. LAEMMLI, U.K.- Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond.), 227:680-685, 1970.
13. NATARO, J.P.; KAPER, J.B.; ROBINS-BROWNE, K. et al. - Pattern of adherence of diarrhoeagenic Escherichia coli to HEp-2 cells. Pediat. infect. Dis. J., 6:829-831, 1987
14. PARRY, S.H. & PORTER, P. - Immunological aspects of cell membrane adhesion demonstrated by porcine enteropathogenic Escherichia coli. Immunology, 34:41-49, 1978.
15. SCALETSKY, I.C.A.; SILVA, M.L.M. & TRABULSI, L.R. -Distinctive patterns of adherence of enteropathogenic Escherichia coli to HeLa cells. Infect. Immun., 45:534-536, 1984.
16. SCALETSKY, I.C.A.; MILANI, S.R.; TRABULSI, L.R. & TRAVASSOS, L. - Isolation and characterization of lhe localized adherence factor of enteropathogenic Echerichia coli. Infect. Immun., 56:2979-2983, 1988.
17. TACKET, C.O.; MANEVAL, D.R. & LEVINE, M.M. - Purification, morphology, and genetics of a new fimbrial putative colonization factor of enterotoxigenic Escherichia coli 0159:H4. Infect. Immun., 55:1063-1069, 1987.
18. YAMAMOTO, T.; WAKISAKA, N.; NAKAE, T. el al. - Characterization of a novel hemagglutinin of diarrhea-associated Escherichia coli that has characteristics of diffusely adhering E. coli and enteroaggregative E. coli. Infect. Immun., 64:3694-3702, 1996.
19. YANO, T.; ARITA, M.; HONDA, T. & MIWATANI, T. - Mannose-resistant hemagglutination factor produced by enteropathogenic Escherichia coli and its role as an adhesive factor. Bact. Adher., 3:99-105, 1989.

Correspondence to:

T. Yano

Caixa Postal 6109

13081-970 Campinas, SP, Brasil

Tel.:(+55-19) 2397945; Fax (+55-19) 2393124

Publication Dates

Publication in this collection
22 Sept 2006
Date of issue
Dec 1996

History

Accepted
13 Dec 1996
Received
23 July 1996

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. BALDINI, M.M.; KAPER, J.B.; LEVINE, M.M. & CANDY, D.C.A. - Plasmid-mediated adhesion in enteropathogenic Escheridia coli. J. pediat. Gastroent. Nutr., 2:534-538, 1983.

[2] 2. BALDINI, M.M.; NATARO, J.P. & KAPER, J.B. - Localization of a determinant for HEp-2 adherence by enterophatogenic Escherichia coli. Infect. Immun., 52:334-336, 1986.

[3] 3. BILGE, S.S.; LAUSEN, C.R.; LAU, W. & MOSELEY, S.L. - Molecular characterization of a fimbrial adhesin, F1845, mediating diffuse adherence of diarrhea-associated Escherichia coli to HEp-2 cells. J. Bact., 171:4281-4289, 1989.

[4] 4. DONNERBERG, M.S. & KAPER, J.B. - Enteropathogenic Escherichia coli. Infect. Immun., 60:3953-3961, 1992.

[5] 5. CATANI, C.F.; YAMADA, A.T.; VIDOTTO, M.C. & YANO, T. -Adhesion of bovine enterotoxigenic Escherichia coli (ETEC) by type l-like fimbriae. FEMS Microbiol. Lett., 137:241-245, 1996.

[6] 6. EVANS, D.G.; EVANS Jr., D.J. & TJOA, W. - Haemagglutination of human group A erythrocythes by enterotoxigenic and enteropathogenic Escherichia coli isolated from adults with diarrhea: correlation with colonization factor. Infect. Immun., 18:330-337, 1977.

[7] 7. GIRON, J.A.; ASY, H.O. & SCHOOLNIK, G.K. - An inducible bundle-forming pilus of enteropathogenic Escherichia coli. Science, 254:710-713, 1991.

[8] 8. GIRON, J.A.; JONES, T.; MILLÁN-VELASCO, F. et al. - Diffuse-adhering Escherichia coli (DAEC) as a putative cause of diarrhea in Mayan children in Mexico. J. infect. Dis., 163:507-513, 1991.

[9] 9. GIRON, J.A.; ASY, H.O. & SCHOOLNIK, G.K. - Characterization of fimbriae produced by enteropathogenic Escherichia coli. J. Bact., 175:7391-7403, 1993.

[10] 10. GOMES, T.A.T.; BLAKE, P.A. & TRABULSI, L.R. - Prevalence of Escherichia coli strains with localized diffuse and aggregative adherence to HeLa cells in infants with diarrhea and matched controls. J. clin. Microbiol., 27:266-269, 1989.

[11] 11. HARTREE, E.F. -Determination of protein. Anal. Biochem., 48:422-427, 1972.

[12] 12. LAEMMLI, U.K.- Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond.), 227:680-685, 1970.

[13] 13. NATARO, J.P.; KAPER, J.B.; ROBINS-BROWNE, K. et al. - Pattern of adherence of diarrhoeagenic Escherichia coli to HEp-2 cells. Pediat. infect. Dis. J., 6:829-831, 1987

[14] 14. PARRY, S.H. & PORTER, P. - Immunological aspects of cell membrane adhesion demonstrated by porcine enteropathogenic Escherichia coli. Immunology, 34:41-49, 1978.

[15] 15. SCALETSKY, I.C.A.; SILVA, M.L.M. & TRABULSI, L.R. -Distinctive patterns of adherence of enteropathogenic Escherichia coli to HeLa cells. Infect. Immun., 45:534-536, 1984.

[16] 16. SCALETSKY, I.C.A.; MILANI, S.R.; TRABULSI, L.R. & TRAVASSOS, L. - Isolation and characterization of lhe localized adherence factor of enteropathogenic Echerichia coli. Infect. Immun., 56:2979-2983, 1988.

[17] 17. TACKET, C.O.; MANEVAL, D.R. & LEVINE, M.M. - Purification, morphology, and genetics of a new fimbrial putative colonization factor of enterotoxigenic Escherichia coli 0159:H4. Infect. Immun., 55:1063-1069, 1987.

[18] 18. YAMAMOTO, T.; WAKISAKA, N.; NAKAE, T. el al. - Characterization of a novel hemagglutinin of diarrhea-associated Escherichia coli that has characteristics of diffusely adhering E. coli and enteroaggregative E. coli. Infect. Immun., 64:3694-3702, 1996.

[19] 19. YANO, T.; ARITA, M.; HONDA, T. & MIWATANI, T. - Mannose-resistant hemagglutination factor produced by enteropathogenic Escherichia coli and its role as an adhesive factor. Bact. Adher., 3:99-105, 1989.

Brasil

Brasil

Purification and parcial characterization of a hemagglutinating factor (HAF): a possible adhesive factor of the diffuse adherent of Escherichia coli (DAEC)

Purificação e caracterização parcial de um fator hemaglutinante (HAF): uma possível adesina de Escherichia coli do tipo aderência difusa (DAEC)

Abstracts

Correspondence to:

Publication Dates

History