Open AccessIdentification of surface-exposed Yersinia pestis proteins by radio-iodination and biotinylation
Author(s) -
Frederico G. C. Abath,
A. M. P. Almeida,
L. C. S. Ferreira
Publication year - 1992
Publication title -
journal of medical microbiology/journal of medical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.91
H-Index - 117
eISSN - 1473-5644
pISSN - 0022-2615
DOI - 10.1099/00222615-37-6-420
Subject(s) - biotinylation , yersinia pestis , avidin , biotin , chemistry , biochemistry , peroxidase , blot , fractionation , virulence , membrane protein , microbiology and biotechnology , biology , enzyme , chromatography , membrane , gene
When whole cells (stationary phase) of Yersinia pestis strain EV76 were radiolabelled with Iodogen and 131I, 16 major and 10 minor surface-exposed outer membrane proteins (OMPs) were identified. Labelling with N-hydroxysuccinimidyl 6-biotinylamino-hexanoate (biotin X-NHS) resulted in a complex protein profile detectable after blotting and developing with peroxidase-conjugated avidin. Y. pestis cell fractionation revealed that biotin X-NHS labelled not only OMPs but also proteins of inner cell compartments. Therefore, radiolabelling was the more reliable technique for identifying the OMPs of Y. pestis.