A glycomic approach reveals a new mycobacterial polysaccharide
File(s)
Accepted version
Author(s)
Type
Journal Article
Abstract
Mycobacterium tuberculosis lipoarabinomannan (LAM) and biosynthetically related lipoglycans and glycans play an important role in host–pathogen interactions. Therefore, the elucidation of the complete biosynthetic pathways of these important molecules is expected to afford novel therapeutic targets. The characterization of biosynthetic enzymes and transporters involved in the formation and localization of these complex macromolecules in the bacterial cell envelope largely relies on genetic manipulation of mycobacteria and subsequent analyses of lipoglycan structural alterations. However, lipoglycans are present in relatively low amounts. Their purification to homogeneity remains tedious and time-consuming. To overcome these issues and to reduce the biomass and time required for lipoglycan purification, we report here the development of a methodology to efficiently purify lipoglycans by sodium deoxycholate–polyacrylamide gel electrophoresis. This faster purification method can be applied on a small amount of mycobacterial cells biomass (10–50 mg), resulting in tens of micrograms of purified lipoglycans. This amount of purified products was found to be sufficient to undertake structural analyses of lipoglycans and glycans carbohydrate domains by a combination of highly sensitive analytical procedures, involving cryoprobe NMR analysis of intact macromolecules and chemical degradations monitored by gas chromatography and capillary electrophoresis. This glycomic approach was successfully applied to the purification and structural characterization of a newly identified polysaccharide, structurally related to LAM, in the model fast-growing species Mycobacterium smegmatis.
Date Issued
2015-08-10
Date Acceptance
2015-08-05
Citation
Glycobiology, 2015, 25 (11), pp.1163-1171
ISSN
1460-2423
Publisher
Oxford University Press (OUP)
Start Page
1163
End Page
1171
Journal / Book Title
Glycobiology
Volume
25
Issue
11
Copyright Statement
© The Author 2015. Published by Oxford University Press. All rights reserved. This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Glycobiology following peer review. The definitive publisher-authenticated version is available online at: http://glycob.oxfordjournals.org/content/25/11/1163
Identifier
http://glycob.oxfordjournals.org/
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
glycomic
Mycobacterium
polysaccharide
BACTERIAL LIPOPOLYSACCHARIDES
GEL-ELECTROPHORESIS
POLYACRYLAMIDE-GELS
MASS-SPECTROMETRY
CELL-ENVELOPE
MOLECULAR-PATTERNS
BOVIS BCG
LIPOARABINOMANNAN
TUBERCULOSIS
SMEGMATIS
Glycomics
Lipopolysaccharides
Mycobacterium tuberculosis
Polysaccharides, Bacterial
11 Medical And Health Sciences
06 Biological Sciences
Publication Status
Published