Mass Spectrometry in the Elucidation of the Glycoproteome of Bacterial Pathogens

Graham, R.L.; Hess, Sonja

doi:10.2174/157016410790979662

Cited by 10 publications

(8 citation statements)

References 213 publications

(368 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For instance, it has been proposed that mannose receptors on host cells might directly interact with mannosylated Mtb proteins to enter the macrophages for survival [2]. While it is now no longer questioned that bacteria including mycobacteria produce glycoproteins [3], our current knowledge about the glycoproteins of Mtb is very limited. In fact, since the first indication that Mtb is reactive towards ConA lectins in 1989 [4], glycosylation sites for only four Mtb glycoproteins have so far been described.…”

Section: Introductionmentioning

confidence: 99%

O-linked glycosylation sites profiling in Mycobacterium tuberculosis culture filtrate proteins

Smith

Sweredoski

Hess

2014

Journal of Proteomics

Self Cite

View full text Add to dashboard Cite

Mycobacterium tuberculosis (Mtb) causes tuberculosis, one of the leading causes of fatal infectious diseases worldwide. Cell-cell recognition between the pathogen Mtb and its host are mediated in part by glycosylated proteins. So far, glycoproteins in Mtb are understudied and for only very few glycoproteins glycosylation sites have been described, e.g., alanine and proline rich secreted protein apa, superoxide dismutase SODC, lipoprotein lpqH and MPB83/MPT83. In this study, glycosylated proteins in Mtb culture filtrate were investigated using liquid chromatography-mass spectrometry approaches and bioinformatic analyses. To validate the presence of glycoproteins, several strategies were pursued including collision induced dissociation, high energy collision dissociation and electron transfer dissociation techniques, and bioinformatics analyses involving a neutral loss search for glycosylated moieties. After extensive data curation, we report glycosylation sites for thirteen Mtb glycoproteins using a combination of mass spectrometry techniques on a dataset collected from culture filtrate proteins. This is the first glycoproteomics study identifying glycosylation sites on mycobacterial culture filtrate proteins (CFP) on a global scale.

show abstract

Section: Introductionmentioning

confidence: 99%

O-linked glycosylation sites profiling in Mycobacterium tuberculosis culture filtrate proteins

Smith

Sweredoski

Hess

2014

Journal of Proteomics

Self Cite

View full text Add to dashboard Cite

show abstract

“…It has been of particular use in the study of the human pathogens, and the elucidation of the Pseudaminic acid, Legionaminic acid and Di-N-Acetyl Bacillosamine biosynthetic pathways in Campylobacter jejuni (reviewed in ref. 103). Matthias Mann and coworkers have investigated the sites of N-glycosylation in two recent papers, 104,105 in which they use an adaptation of the 'Filter Aided Sample Preparation' (FASP) protocol 106 termed N-Glyco-FASP.…”

Section: Identifying Post-translation Modification (Ptm) Of Proteinsmentioning

confidence: 99%

Proteomics in the microbial sciences

Graham¹,

McMullan²,

Graham³

2011

Bioengineered Bugs

View full text Add to dashboard Cite

Mass spectrometry based proteomics is now widely used in the microbial sciences. In conjunction with transcriptomics it has greatly enhanced the field of microbial biology and has provide microbiologists with unparalleled insights into cellular processes and functions. Proteomics allows the dynamic nature of the entire protein network to be mapped providing a deeper understanding of microbial systems, their evolution and role in disease states. This review is intended to provide an overview of mass spectrometry and its application to the field of microbial proteomics. Background is provided on the core mass analyzers, including the Orbitrap mass spectrometer, and novel fragmentation processes such as Electron Transfer Dissociation which leave post-translational modifications intact on peptide backbones allowing for their identification and localization. The review will also provide information on current key quantitative technologies and the state of the art in microbial metaproteomics.

show abstract

“…It is presumed to be the result of an additional hydrolase activity of PglB [7] because the assembly of the glycan to the protein is done in consecutive steps, one sugar unit after another [8,1]. Thus, free oligosaccharides (fOS) would not be expected to be present from the assembly.…”

Section: Introductionmentioning

confidence: 99%

“…Zwitterionic hydrophilic interaction chromatography (ZIC-HILIC), known to have high capability to separate eukaryotic N-glycans or glycopeptides [10,11], has been established for the studies in Campylobacter species [9,8]. The enriched glycans and glycoprotein can then be analyzed by mass spectrometry.…”

Section: Introductionmentioning

confidence: 99%

A cationic cysteine-hydrazide as an enrichment tool for the mass spectrometric characterization of bacterial free oligosaccharides

et al. 2015

Self Cite

View full text Add to dashboard Cite

In Campylobacterales and related ε-proteobacteria with N-linked glycosylation (NLG) pathways, free oligosaccharides (fOS) are released into the periplasmic space from lipid-linked precursors by the bacterial oligosaccharyltransferase (PglB). This hydrolysis results in the same molecular structure as the oligosaccharide that is transferred to a protein to be glycosylated. This allowed for the general elucidation of the fOS branched structures and monosaccharides from a number of species using standard enrichment and mass spectrometry methods. To aid characterization of fOS, hydrazide chemistry has often been used for chemical modification of the reducing part of oligosaccharides resulting in better selectivity and sensitivity in mass spectrometry; however, the removal of the unreacted reagents used for the modification often causes the loss of the sample. Here, we develop a more robust method for fOS purification and characterize glycostructures using complementary tandem mass spectrometry (MS/MS) analysis. A cationic cysteine hydrazide derivative was synthesized to selectively isolate fOS from periplasmic fractions of bacteria. The cysteine hydrazide nicotinamide (Cyhn) probe possesses both thiol and cationic moieties. The former enables reversible conjugation to a thiol-activated solid support, while the latter improves the ionization signal during MS analysis. This enrichment was validated on the well-studied Campylobacter jejuni by identifying fOS from the periplasmic extracts. Using complementary MS/MS analysis we approximated data of a known structure of the fOS from Campylobacter concisus. This versatile enrichment technique allows for the exploration of a diversity of protein glycosylation pathways.

show abstract

Mass Spectrometry in the Elucidation of the Glycoproteome of Bacterial Pathogens

Cited by 10 publications

References 213 publications

O-linked glycosylation sites profiling in Mycobacterium tuberculosis culture filtrate proteins

O-linked glycosylation sites profiling in Mycobacterium tuberculosis culture filtrate proteins

Proteomics in the microbial sciences

A cationic cysteine-hydrazide as an enrichment tool for the mass spectrometric characterization of bacterial free oligosaccharides

Contact Info

Product

Resources

About