Bacterial glycoproteomics

Abstract

Glycosylated proteins are ubiquitous components of eukaryote cellular surfaces, where the glycan moieties are implicated in a wide range of cell–cell recognition events. Once thought to be restricted to eukaryotes, glycosylation is now being increasingly reported in prokaryotes. Many of these discoveries have grown from advances in analytical technologies and genome sequencing. This review highlights the capabilities of high-sensitivity mass spectrometry for carbohydrate structure determination of bacterial glycoproteins and the emergence of glycoproteomic strategies that have evolved from proteomics and genomics for the functional analysis of bacterial glycosylation.

Conservation of the PTEN catalytic motif in the bacterial undecaprenyl pyrophosphate phosphatase, BacA/UppP

“…In addition, undecaprenyl monophosphate is the carrier for N-linked protein glycosylation in prokaryotes (Fig. 1b) (Hitchen & Dell, 2006;Kumar & Balaji, 2011;Tabish et al, 2011). Smaller polyisoprenoids such as decaprenyl phosphate (C 50 -P) are also as carriers for lipid I and II in mycobacteria (Mahapatra et al, 2005).…”

“…Bacterial monophosphate lipid carriers (Hitchen & Dell, 2006;Kumar & Balaji, 2011;Nothaft & Szymanski, 2010) and dolichyl pyrophosphate (Dol-PP), through a GlcNAc linkage, is complexed with a high mannose tree involved in eukaryotic protein glycosylation (Fig. S1C) (Cantagrel & Lefeber, 2011;Helenius & Aebi, 2004;Larkin & Imperiali, 2011;Schwarz & Aebi, 2011;Welti, 2013).…”

“…Each of these lipid carriers must first exist in the monophosphate state to carry out their respective roles. As part of the transfer of the cargo to their lipid carriers, the polyisoprenyl lipid monophosphate is converted to a pyrophosphate containing their respective cargos (Jones et al, 2009): undecaprenyl pyrophosphate carries MurNAc-GlcNAc-peptide complexes for bacterial cell envelope biosynthesis (Figs 1b and S1A, available in Microbiology Online) (Bouhss et al, 2008;Hitchen & Dell, 2006;Lovering et al, 2012), undecaprenyl pyrophosphate binds a heptasaccharide for N-linked glycosylation in bacteria as identified in Campylobacter jejuni ( (a) Dol-P is used as the lipid carrier for eukaryotic protein glycosylation. N-Acetylglucosamine and mannose are added to Dol-P in the cytoplasm, which is then believed to be flipped to the ER lumen.…”

Glycoproteomics: Past, present and future

“…For example, although not always applicable to large and/or highly heterogeneous glycoproteins, molecular weight profiling of intact glycoproteins (purple arrows in Figure 1) can sometimes provide very useful information on the type and extent of glycosylation. Such "top-down" methods have proven especially powerful, in bacterial glycoproteomics where novel glycans are frequently observed [26][27][28], and in studies on intact antibodies for the Biopharmaceutical industry, where M-Scan routinely screens intact masses at around 150 kilodaltons by both MALDI-TOF and ES-Q-TOF to give confirmatory total mass analysis when reconstructing the detailed protein and carbohydrate profiles from mass mapping studies.…”

Application of proteomics to ecology and population biology

“…The application of MS-based techniques for the qualitative and quantitative analysis of cellular proteomes, often from complex mixtures, has become an important tool for the understanding of cellular function across a wide diversity of plant and animal taxa (Fels et al, 2003;Han and Lee, 2003;Ahram and Springer, 2004;Aldred et al, 2004;Banta-Wright and Steiner, 2004;Chen and White, 2004;Baginsky and Gruissem, 2006;Berven et al, 2006;Bowler et al, 2006;Hitchen and Dell, 2006) Historically, MS has been a workhorse analytic tool used by chemists and associated industries, where it is mainly used as a tool for structural determination of organic compounds and quality control assurance in the pharmacetutical and biotech industries. MS is also a prominent tool used by security agencies, for example in rapid detection of explosive materials at airports (Takats et al, 2004(Takats et al, , 2005a Over the past two decades, a number of major advances in MS technologies applied to the analysis of proteins and peptides has elevated MS to a central role in proteomics, including new methods for the introduction of complex protein and peptide mixtures into the mass spectrometer in such a manner as to allow for the de novo sequencing of small peptides (Lin et al, 2003;Yates, 2004;Gingras et al, 2005;Guerrera and Kleiner, 2005;Domon and Aebersold, 2006) Combined with the ever-expanding availability of whole, or partial, genome sequences and bioinformatic tools for their analysis, MS has assumed a primary role in proteomic analyses.…”