title: Cell wall proteome of Clostridium thermocellum and detection of glycoproteins.
abstract: Clostridium thermocellum, a thermophilic anaerobe, has the unusual capacity to convert cellulosic biomass into ethanol and hydrogen. In this work, the cell wall proteome of C. thermocellum was investigated. The proteins in the cell wall fraction of C. thermocellum prepared by the boiling SDS method were released by mutanolysin digestion and resolved on two-dimensional (2D) gel. One hundred and thirty-two proteins were identified by mass spectrometry, among which the extracellular solute-binding protein (CbpB/cthe_1020), enolase, glyceraldehyde-3-phosphate dehydrogenase and translation elongation factor EF-Tu were detected as highly abundant proteins. Besides the known surface localized proteins, including FtsZ, MinD, GroEL, DnaK, many enzymes involved in bioenergetics, such as alcohol dehydrogenases and hydrogenases were also detected. By glycan stain and MS analysis of glycopeptides, we identified CbpB as a glycoprotein, which is the second glycoprotein from C. thermocellum characterized. The fact that CbpB was highly abundant in the cell wall region and glycosylated, reflects its importance in substrate assimilation. Our results indicate cell wall proteins constitute a significant portion of cellular proteins and may play important physiological roles (i.e. bioenergetics) in this bacterium. The insights described are relevant for the development of C. thermocellum as a biofuel producer.
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Yu
authors.forename: Xinping
authors.initials: X
authors.lastname: Xu
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Peng
authors.forename: Yuanming
authors.initials: Y
authors.lastname: Luo
authors.forename: Keqian
authors.initials: K
authors.lastname: Yang
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis*; D002473:Cell Wall / Q000737:chemistry*; D048013:Acetivibrio thermocellus / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D006023:Glycoproteins / Q000032:analysis*; D013058:Mass Spectrometry; D011134:Polysaccharides / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D006023:Glycoproteins; D011134:Polysaccharides; D020543:Proteome