Sialylation of lipopolysaccharide by CMP-NANA in viable gonococci is enhanced by low Mr material released from blood cell extracts but not by some UDP sugars.
Serum resistance of gonococci in most patients is due to sialylation of a Gal beta 1-4GlcNAc group on a conserved 4.5 kDa lipopolysaccharide (LPS) component by host cytidine 5'-monophospho-N-acetyl neuraminic acid (CMP-NANA) catalysed by a gonococcal sialyl transferase. This sialylation is enhanced by a low M(r) factor(s) which, like CMP-NANA, is released in diffusates from high M(r) fractions obtained from sonicates dialysed at 4 degrees C. Also, as shown here, this factor(s) is released when the sonicates are dialysed at 18-20 degrees C. The enhancement of sialylation, first demonstrated using enzymes in gonococcal extracts, has been shown to occur in live gonococci and hence probably to have a role in pathogenicity. Gonococci, emerging from lag phase and incubated for 2 h with CMP-14CNANA fixed up to 90% more radiolabel than controls when the second factor(s) was present; their LPS separated by SDS-PAGE contained more radiolabel than control samples and label was not detected in any other component. Fractions with enhancing activity absorbed maximally at about 260 nm but a mixture of UDP-galactose (UDP-Gal), UDP-N-Acetyl galactosamine (UDP-GalNAc), UDP-glucose (UDP-Glc) and UDP-N-Acetyl glucosamine (UDP-GlcNAc) showed no significant enhancing activity. The enhancing action of the low M(r) fractions was unaffected by incubation with beta-galactosidase.
Parsons
N J
NJ
School of Biochemistry, University of Birmingham, U.K.
Constantinidou
C
C
Cole
J A
JA
Smith
H
H
D001770:Blood Bactericidal Activity; D002240:Carbohydrate Sequence; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid / Q000378:metabolism*; D006069:Gonorrhea / Q000097:blood / Q000382:microbiology; D006801:Humans; D066298:In Vitro Techniques; D008070:Lipopolysaccharides / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D012794:Sialic Acids / Q000737:chemistry / Q000378:metabolism; D014539:Uridine Diphosphate Sugars / Q000494:pharmacology
D008070:Lipopolysaccharides; D012794:Sialic Acids; D014539:Uridine Diphosphate Sugars; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid