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Chembiochem ; 16(6): 977-89, 2015 Apr 13.
Artículo en Inglés | MEDLINE | ID: mdl-25766777

RESUMEN

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a major pathogen responsible for 1.5 million deaths annually. This bacterium is characterized by a highly unusual and impermeable cell envelope, which plays a key role in mycobacterial survival and virulence. Although many studies have focused on the composition and functioning of the mycobacterial cell envelope, the capsular α-glucan has received relatively minor attention. Here we show that a murine monoclonal antibody (Mab) directed against glycogen cross-reacts with mycobacterial α-glucans, polymers of α(1-4)-linked glucose residues with α(1-6)-branch points. We identified the Mab epitope specificity by saturation transfer difference NMR and show that the α(1-4)-linked glucose residues are important in glucan-Mab interaction. The minimal epitope is formed by (linear) maltotriose. Notably, a Mycobacterium mutant lacking the branching enzyme GlgB does not react with the Mab; this suggests that the α(1-6)-branches form part of the epitope. These seemingly conflicting data can be explained by the fact that in the mutant the linear form of the α-glucan (amylose) is insoluble. This Mab was subsequently used to develop several techniques helpful in capsular α-glucan research. By using a capsular glucan-screening methodology based on this Mab we were able to identify several unknown genes involved in capsular α-glucan biogenesis. Additionally, we developed two methods for the detection of capsular α-glucan levels. This study therefore opens new ways to study capsular α-glucan and to identify possible targets for further research.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Cápsulas Bacterianas/metabolismo , Epítopos/inmunología , Glucógeno/inmunología , Glucógeno/metabolismo , Mycobacterium/metabolismo , Animales , Pared Celular/metabolismo , Elementos Transponibles de ADN/genética , Glucógeno/biosíntesis , Glucógeno/química , Espectroscopía de Resonancia Magnética , Ratones , Mutación , Mycobacterium/citología , Oligosacáridos/química
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