Systematic Characterization of Protein Glycosylation of Bacteria Cell Surface Proteins
EMSL Project ID
45394
Abstract
The overall goal of this proposal is to develop an integrated top-down and bottom-up approach for bacterial glycoprotein profiling assisted by accurate glycan structure identification, which will be applied to study bacterial glycosylations. This platform will combine multiple cutting-edge "omics" techniques (proteomics, glycomics, and bioinformatics) uniquely provided by EMSL: (1) state-of-the-art instrumentation (FTMS, including high field FTICR MS, high resolution NMR); (2) versatile in-house built high performance LC platforms, which provide unparalleled throughput and sensitivity for both intact proteins and peptides; (3) computing capabilities and in-house software developments; and (4) staff expertise in the areas of MS, HPLC, NMR and bioinformatics.
Using the proposed platform, we will test the hypothesis that the structure of the glycan moieties of thermophilic bacteria glycoproteins are different from those of bacteria that thrive at low temperatures. To accomplish this, we will perform the following tasks:
1. Apply our proposed platform to profile the glycoproteome in Synechococcus sp. strain PCC 7002, a model cyanobacteria system for which limited information on glycosylations is already available.;
2. Apply our proposed platform to find the glycoproteome signature in extremely thermophilic bacteria, T. maritima and K. olearia.
Project Details
Start Date
2011-07-11
End Date
2014-07-20
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Cao L, N Tolic, Y Qu, D Meng, R Zhao, Q Zhang, RJ Moore, EM Zink, MS Lipton, L Pasa-Tolic, and S Wu. 2014. "Characterization of intact N-linked and O-linked glycopeptides using higher energy collisional dissociation ." Analytical Biochemistry. doi:10.1016/j.ab.2014.01.003
Hengel SM, S Wu, EA Floyd, ES Baker, R Zhao, and L Pasa-Tolic. 2012. "Evaluation of SDS depletion using an affinity spin column and IMS-MS detection." PNNL-SA-87924, Pacific Northwest National Laboratory, Richland, WA.
Liu X, SM Hengel, S Wu, N Tolic, L Pasa-Tolic, and PA Pevzner. 2013. "Identification of Ultramodified Proteins Using Top-Down Mass Spectra." PNNL-SA-94025, Pacific Northwest National Laboratory, Richland, WA. doi:10.1007/978-3-642-37195-0_11 [Unpublished]
Meng D, Q Zhang, X Gao, S Wu, and G Lin. 2014. "LipidMiner: a software for automated identification and quantification of lipids from multiple liquid chromatography-mass spectrometry data files." Rapid Communications in Mass Spectrometry 28(8):981-985. doi:10.1002/rcm.6865
Payne SH, S Bonissone, S Wu, RN Brown, D Ivankov, D Frishman, L Pasa-Tolic, RD Smith, and PA Pevzner. 2012. "Unexpected Diversity of Signal Peptides in Prokaryotes." mBio 3(6):Article No. e00339-12. doi:10.1128/mBio.00339-12.
Qu Y, S Wu, R Zhao, EM Zink, DJ Orton, RJ Moore, D Meng, TRW Clauss, JT Aldrich, MS Lipton, and L Pasa-Tolic. 2013. "Automated Immobilized Metal Affinity Chromatography System for Enrichment of Escherichia coli Phosphoproteome." Electrophoresis 34(11):1619-1626. doi:10.1002/elps.201200628
Wu S, RN Brown, SH Payne, D Meng, R Zhao, N Tolic, L Cao, AK Shukla, ME Monroe, RJ Moore, MS Lipton, and L Pasa-Tolic. 2013. "Top-down characterization of the post-translationally modified intact periplasmic proteome of the bacterium Novosphingobium aromaticivorans ." International Journal of Proteomics 2013:279590. doi:10.1155/2013/279590.