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Revista do Instituto de Medicina Tropical de São Paulo

On-line version ISSN 1678-9946

Rev. Inst. Med. trop. S. Paulo vol.32 no.2 São Paulo Mar./Apr. 1990

http://dx.doi.org/10.1590/S0036-46651990000200002 

ORIGINAL ARTICLES

 

Comparative studies of Yersinia pestis outer membrane isolation techniques and their potential use in plague epidemiology

 

Estudo comparativo de técnicas de isolamento de membrana externa de Yersinia pestis e seu uso na epidemiologia da peste

 

 

Frederico Guilherme Coutinho AbathI; Luís Carlos de Sousa FerreiraII

ICentro de Pesquisas Aggeu Magalhães (Fundação Oswaldo Cruz). Caixa Postal 7472. CEP 59.739 Recife, Pernambuco, Brazil
IILaboratório de Imunopatologia Keizo Asami e Departamento de Biofísica e Radiobiologia da Universidade Federal de Pernambuco. CEP 59739 Recife, Pernambuco, Brazil

Present address

 

 


SUMMARY

In the present study three techniques for obtaining outer membrane enriched fractions from Yersinia pestis were evaluated. The techniques analysed were: differential solubilization of the cytoplasmic membrane with Sarkosyl or Triton X-100, and centrifugation in sucrose density gradients. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of outer membrane isolated by the different methods resulted in similar protein patterns. The measurement of NADH-dehydrogenase and succinate dehydrogenase (inner membrane enzymes) indicated that the outer membrane preparations obtained by the three methods were pure enough for analytical studies. In addition, preliminary evidences on the potential use of outer membrane proteins for the identification of geographic variants of Y. pestis wild isolates are presented.

Key words: Yersinia pestis, outer membrane proteins, membrane isolation techniques.


RESUMO

No presente estudo três técnicas para isolamento de frações enriquecidas em membrana externa de Y. pestis foram avaliadas. As técnicas utilizadas foram: centrifugação em gradiente de densidade em sacarose e solubilização diferencial com Sarkosyl ou Triton X-100. A análise por eletroforese em gel de poliacrilamida na presença de dodecil sulfato de sódio (SDS-PAGE) das membranas externas extraídas pelos diferentes métodos evidenciou perfis protéicos semelhantes. A determinação das atividades de NADH-desidrogenase e succinato-desidrogenase (enzimas de membrana interna) indicou que todas as preparações estudadas eram adequadas a estudos analíticos. Obteve-se evidências preliminares sobre o possível uso de perfis protéicos de membrana externa na identificação de variantes geográficos entre isolados selvagens de Y. pestis.


 

 

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

 

 

ACKNOWLEDGMENTS

We are acknowledged to Dra. A. M. P. Almeida for helpful, advice, incentive and bacterial strains; Dr. R. R. Brubaker for the kind gift of strains A1122 and KIM and Mr. H. S. Almeida and Mrs. Y. Nakasawa for the helpful technical assistance. Research supported by the Japanese Cooperation Agency.

 

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Present address:
Dr. Luiz Carlos de Souza Ferreira
Universidade Federal do Rio de Janeiro, Instituto de Biofísica Carlos Chagas Filho
CEP 21.941 Rio de Janeiro, Brasil

Recebido para publicação em 18/5/1989.

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