New structural insights into the molecular deciphering of mycobacterial lipoglycan binding to C-type lectins: lipoarabinomannan glycoform characterization and quantification by capillary electrophoresis at the subnanomole level.
Lipoarabinomannans are key molecules of the mycobacterial envelopes involved in many steps of tuberculosis immunopathogenesis. Several of the biological activities of lipoarabinomannans are mediated by their ability to bind human C-type lectins, such as the macrophage mannose receptor, the mannose-binding protein and the surfactant proteins A and D. The lipoarabinomannan mannooligosaccharide caps have been demonstrated to be involved in the binding to the lectin carbohydrate recognition domains. We report an original analytical approach, based on capillary electrophoresis monitored by laser-induced fluorescence, allowing the absolute quantification, in nanomole quantities of lipoarabinomannan, of the number of mannooligosaccharide units per lipoarabinomannan molecule. Moreover, this analytical approach was successful for the glycosidic linkage determination of the mannooligosaccharide motifs and has been applied to the comparative analysis of parietal and cellular lipoarabinomannans of Mycobacterium bovis BCG and Mycobacterium tuberculosis H37Rv, H37Ra and Erdman strains. Significant differences were observed in the amounts of the various mannooligosaccharide units between lipoarabinomannans of different strains and between parietal and cellular lipoarabinomannans of the same strain. Nevertheless, no relationship was found between the number of mannooligosaccharide caps and the virulence of the corresponding strain. The results of the present study should help us to gain more understanding of the molecular basis of lipoarabinomannan discrimination in the process of binding to C-type lectins.
Nigou
J
J
Institut de Pharmacologie et de Biologie Structurale, Centre National de la Recherche Scientifique, UPR 9062, 205 route de Narbonne, Toulouse Cedex 4, 31077, France.
Vercellone
A
A
Puzo
G
G
D000107:Acetylation; D000942:Antigens, Bacterial / Q000032:analysis / Q000737:chemistry; D002236:Carbohydrate Conformation; D019075:Electrophoresis, Capillary / Q000379:methods; D005453:Fluorescence; D006801:Humans; D007834:Lasers; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008351:Mannans / Q000032:analysis / Q000737:chemistry; D009163:Mycobacterium bovis / Q000737:chemistry* / Q000145:classification / Q000472:pathogenicity; D009169:Mycobacterium tuberculosis / Q000737:chemistry* / Q000145:classification / Q000472:pathogenicity; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014774:Virulence
D000942:Antigens, Bacterial; D037102:Lectins; D008070:Lipopolysaccharides; D008351:Mannans; D009844:Oligosaccharides; C050016:lipoarabinomannan