Identification of surface-exposed Yersinia pestis proteins by radio-iodination and biotinylation.
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.
Abath
F G
FG
Laboratório de Immunopatologia Keizo Asami, Universidade Federal de Pernambuco, Recife, Brazil.
Almeida
A M
AM
Ferreira
L C
LC
D001425:Bacterial Outer Membrane Proteins / Q000032:analysis*; D001710:Biotin / Q000031:analogs & derivatives; D002458:Cell Fractionation; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D013388:Succinimides; D014508:Urea / Q000031:analogs & derivatives; D015010:Yersinia pestis / Q000737:chemistry*
D001425:Bacterial Outer Membrane Proteins; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; C078636:N-hydroxysuccinimidyl 6-biotinylaminohexanoate; D013388:Succinimides; C012385:1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril; D001710:Biotin; D014508:Urea