In vivo determination of Neisseria meningitidis serogroup A capsular polysaccharide by whole cell high-resolution magic angle spinning NMR spectroscopy.
High resolution-magic angle spinning (HRMAS) NMR spectroscopy was applied to serogroup A Neisseria meningitidis (NMA) to determine precise structures of capsular polysaccharide (CPS) expressed on the meningococcal surface. Both the O-acetylated (OAc) NMA parent and a mynC::aphA3 OAc- mutant demonstrated characteristic CPS-derived NMR signals indicating cell-surface expression of CPS, but only the parent expressed O-3 and O-4 acetylation signals. A capsule-defective strain showed no NMR signals for CPS. The (1)H NMR HRMAS spectral patterns correlated with the purified CPS (1)H NMR profiles. HRMAS NMR can distinguish detailed complex carbohydrate structures expressed on bacteria. NMA express both O-3 and O-4 acetylated polymers but not in equimolar ratio amounts in vivo.
Gudlavalleti
Seshu K
SK
Laboratory of Microbial Pathogenesis, VA Medical Center and Division of Infectious Diseases, Atlanta, GA, USA.
Szymanski
Christine M
CM
Jarrell
Harold C
HC
Stephens
David S
DS
D016667:Bacterial Capsules / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D038381:Neisseria meningitidis, Serogroup A / Q000737:chemistry*; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D013499:Surface Properties
D011135:Polysaccharides, Bacterial