For the recent 20 years I've been working in the N.D. Zelinsky Institute of Organic Chemistry NMR center as glycopolymer structural researcher in the group of Prof. Knirel (partly as a phd student supervised by Prof. Shashkov). Our systematic chemical and immunochemical studies are aimed at correlation between the O-antigen structure and the immunospecificity of strains, which is important for obtaining synthetic diagnosticums, vaccines and immunostimulators.
Besedes a bacterial class on which I focused in 1996-2002 (Proteus studies), a lot of other bacterial O-antigens were investigated (see below).
The other projects whichg I participated or carried out personally:
Glyco-studies included the elucidation of structure of natural glycopolymers and glycoconjugates by NMR spectroscopic methods. Click here to view the approximate plan of a glycopolymer NMR investigation. The NMR experiments were carried out on Bruker NMR instruments (WM250, AM300, DRX500) and included the following:
|NMR experiment||Its result|
|1D 13C NMR||preliminary data, glycosylation effects (=> substitution pattern, absolute configurations)|
|1D 1H NMR||HH-coupling constants analysis (=> monomeric composition, anomeric configurations)|
|1D 31P NMR||revealing non-sugar substituents|
|1D H-Gated 13C NMR||anomeric configuration, cycle sizes|
|1D Attached Proton Test||methylen groups assignment|
|2D 1H,1H COSY||vicinal proton-proton interactions (=> 1H NMR spectrum assignment)|
|2D 1H,1H Double Quantum Filter COSY||simplifying the COSY spectrum by the removal of diagonal line|
|2D H,H-relayed COSY (RCT, RCT2)||far proton-proton interactions (=> 1H NMR spectrum assignment)|
|2D TOtal Correlation SpectroscopY||excluding the spin system subsets (of residues) from 1H NMR spectrum|
|1D 1H difference mode double resonance||signal shape analysis (=> thorough 1H NMR spectrum assignment )|
|2D 31P,1H COSY||localization of phospho-containing substituents|
|2D 13C,1H COSY||direct heteronuclear coupling constants (=> 13C NMR spectrum assignment)|
|2D 1H,13C Heteronuclear Multiple-Quantum Coherence||direct heteronuclear coupling constants (=> 13C NMR spectrum assignment, constant measurement)|
|2D Heteronuclear Multiple-Bond Correlation||far heteronuclear interactions, including transglycosidic (=> residue sequence)|
|2D NOE SpectroscopY||revealing spatial H-H contacts in oligomers, including transglycosydic (=> substitution pattern, residue sequence)|
|2D Rotating frame nuclear Overhauser Effect SpectroscopY||revealing spatial H-H contacts, including transglycosydic (=> substitution pattern, residue sequence)|
|1D difference mode NOE spectroscopy||thorough analysis of H-H spatial contacts|
Structural studies of the Proteus O-antigens were supported by more than 10 RFBR and INTAS grants (grant list here).