Preparation and characterisation of envelope proteins from Haemophilus pleuropneumoniae.
Envelope proteins of Haemophilus pleuropneumoniae were extracted by 3 methods and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Three major envelope proteins (45,000 Mr, 41,000 Mr, 31,500 Mr) were distinguished in sonicated cell envelopes together with minor proteins. Using selective solubilisation with sodium lauryl sarcosinate or Triton X-100, outer membrane proteins were distinguished from those of the cytoplasmic membrane. Extraction into LiCl produced a similar profile, but the 41,000 Mr and 31,500 Mr bands were present in reduced amounts. Extraction into saline at 60 degrees C produced a grossly different pattern, with a major band at 20,000 Mr. All 3 major envelope proteins were shown to be heat-modifiable, and the 31,500 Mr band was found to be the non-heat-modified form of a 43,000 Mr protein, which showed similar properties to the Protein d of H. influenzae which is related to the OmpA protein of E. coli K-12. The 45,000 Mr major protein was also weakly associated with the peptidoglycan in SDS/Triton at low temperature.
Rycroft
A N
AN
Department of Veterinary Pathology, University of Glasgow Veterinary School, Gt. Britain.
Taylor
D J
DJ
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000032:analysis* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006190:Haemophilus / Q000032:analysis*; D006358:Hot Temperature; D013552:Swine
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins