FIG. 5.

AniA and GNA1946 are glycosylated in MC58. (A) Equivalent whole-cell lysate samples of N. meningitidis MC58 were electrophoresed in adjacent lanes, and following transfer to a PVDF membrane, the membrane was divided in half lengthwise. The two pieces were then probed with MAb npg2 and antibodies to AniA. The results demonstrate that the same band corresponding to AniA was identified with both antibodies. (B) Identification of the glycopeptide ⁶DSAPAASASAAADNGAAK²³ from lipoprotein GNA1946 derived by tryptic cleavage of a membrane protein-enriched fraction of MC58. High-energy collision dissociation (HCD) fragmentation of [M+2H]²⁺ at m/z 989.4505 revealed characteristic oxonium ions at m/z 433.182 and m/z 229.118 corresponding to O-Ac-Hex-DATDH and DATDH glycans, respectively. The signal at m/z 1545.716 is a signal from the peptide backbone after loss of the glycan moiety; the monoisotopic theoretical molecular mass is 1,545.714 Da.