Covalent linkage between the capsular polysaccharide and the cell wall peptidoglycan of Streptococcus pneumoniae revealed by immunochemical methods.
The attachment of capsular polysaccharide to Streptococcus pneumoniae was examined using monoclonal and polyclonal antibodies. Among the strains examined, the capsular polysaccharide of types 2, 4, 6A, 6B, 7F, 8, 14, 19F and 23F was bound to the pneumococci whereas that of a type 3 strain was not. Sequential treatment with 2% SDS at 100 degrees C, pronase, and EDTA did not dissociate the capsular polysaccharide from the pneumococci. Treatment of the cells with mutanolysin, a muramidase that degrades the cell wall peptidoglycan of pneumococci and other streptococci, released both the capsular and the cell wall C-polysaccharide (C-Ps). Type 6A capsular polysaccharide released from cell walls by mutanolysin treatment, was fractionated by high performance liquid chromatography and examined by immunoelectrophoresis. It was found to be bound to both the C-Ps and the peptidoglycan. The bond between the capsular polysaccharide and the peptidoglycan has not yet been identified but is probably covalent, as the two components could not be dissociated after boiling in SDS. Based on our studies with type 6A, we propose that capsular polysaccharide and C-Ps of the pneumococcus are linked to the peptidoglycan at different sites and, thereby, indirectly to each other. Studies in mice showed that the peptidoglycan enhanced the serum antibody response to C-Ps but not to type 6A polysaccharide.
Sørensen
U B
UB
Laboratory of Developmental and Molecular Immunity, National Institute of Health and Human Development, NIH, Bethesda, Maryland 20892.
Henrichsen
J
J
Chen
H C
HC
Szu
S C
SC
D000596:Amino Acids / Q000032:analysis; D002473:Cell Wall / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D010450:Endopeptidases / Q000378:metabolism; D007122:Immunoelectrophoresis; D010457:Peptidoglycan / Q000378:metabolism*; D011135:Polysaccharides, Bacterial / Q000032:analysis / Q000378:metabolism*; D013296:Streptococcus pneumoniae / Q000032:analysis / Q000378:metabolism*
D000596:Amino Acids; D010457:Peptidoglycan; D011135:Polysaccharides, Bacterial; C027158:polysaccharide C-substance (Streptococcus); D010450:Endopeptidases; C021816:mutanolysin