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)

Yano Tomomasa Cleide Ferreira Catani Michiko Arita Takeshi Honda Toshio Miwatani About the authors

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 YanoI; Cleide Ferreira CataniI; Michiko AritaII; Takeshi HondaII; Toshio MiwataniII

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

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.

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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).

  • Correspondence to:
    T. Yano
    Caixa Postal 6109
    13081-970 Campinas, SP, Brasil
    Tel.:(+55-19) 2397945; Fax (+55-19) 2393124
  • Recebido para publicação em 23/07/1996

    Aceito para publicação em 13/12/1996

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    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
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