Analysis of pneumococcal PspA microheterogeneity in SDS polyacrylamide gels and the association of PspA with the cell membrane.
Pneumococcal surface protein A (PspA) is a protection-eliciting surface protein found on all pneumococci. Although highly cross-reactive, it displays interstrain variation in its size and in the expression of individual antibody reactive epitopes. PspA was not released in significant amounts from pneumococcal membranes treated with sodium carbonate, but was solubilized with SDS. Thus, PspA is either an integral membrane protein or is attached to an integral membrane component. By SDS-PAGE and immunoblot analysis, we found two predominant molecular sizes of PspA in each strain examined. The smaller band was about the size expected from the inferred amino acid sequence of PspA and the larger band appeared to be a dimer of the monomer PspA. When higher concentrations of lysate were run on SDS gels, it was also possible to detect many additional high molecular weight components that reacted with antibodies to PspA. These multiple high molecular weight PspA bands were not due to the attachment of PspA to peptidoglycan or teichoic acids, did not appear to be composed of degraded PspA and most likely resulted from non-covalent polymerization or aggregation of PspA.
Talkington
D F
DF
Department of Microbiology, University of Alabama, Birmingham 35294.
Voellinger
D C
DC
McDaniel
L S
LS
Briles
D E
DE
D000954:Antigens, Surface / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010457:Peptidoglycan / Q000737:chemistry; D013296:Streptococcus pneumoniae / Q000737:chemistry*; D013682:Teichoic Acids / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry
D000954:Antigens, Surface; D001426:Bacterial Proteins; D010457:Peptidoglycan; D013682:Teichoic Acids; C051592:pneumococcal surface protein A; D014357:Trypsin