RJPB, MI and GC performed
the bioinformatic analysis and participated in genome comparison. MDG and FI participated S63845 molecular weight in the analysis and comparison of the exogenous genetic elements. ER performed DNA preparation and generated the 454 sequencing data. FS and MM carried out the ultrastructural characterization of phage particles. LM participated in the genome comparison. GDB participated in the design of the study, its coordination and helped in revising the manuscript. MRO participated in the design of the study, carried out the genome comparison and helped in writing the manuscript. AP participated in the design of the study, its coordination and finalized the manuscript. All authors read and approved the final manuscript.”
“Background Clostridium thermocellum is a Gram-positive thermophilic anaerobe capable of degrading cellulose and producing ethanol and hydrogen. These qualities render C. thermocellum potentially useful for the production of biofuel from biomass. The cellulytic activities of this organism were well selleck inhibitor studied, the corresponding enzymes were found to organize into a cell surfaced bound multienzyme complex, termed cellulosome . The arrangement of the enzymatic subunits in the cellulosome complex, made possible by a scaffoldin subunit, promotes
enhanced substrate binding and degradation. However, other parts of its cellular functions are not well understood. Recently, a genome scale metabolic model was constructed , which provides a good basis for the overall understanding of its metabolism. Since membrane is where many important physiological functions, such as energy generation, protein trafficking, and small molecule transport , take place, we focused on membrane protein complexes as a start point to identify unique features of C. thermocellum. Identification of
protein complexes in C. thermocellum is an important step toward understanding cellular behavior at an integrative level. Blue native-PAGE Tacrolimus (FK506) (BN-PAGE) is a charge shift method first developed by Schägger and von Jagow  to separate membrane protein complexes. It has been used successfully to characterize respiratory complexes in yeast mitochondria and Paracoccus denitrificans [5, 6], photosynthetic complexes in plants and Synechocystis [7, 8], and cell envelope protein complexes in E. coli [9, 10]. It differs from other native gel electrophoresis mainly because the electrophoretic mobility of a protein is determined by the negative charge of the bound Coomassie blue dye, while separation of proteins is achieved by the molecular sieve effect provided by the polyacrylamide gradient of ZD1839 descending pore size similar to other PAGE methods. BN-PAGE, when coupled with a second dimensional SDS-PAGE and mass spectrometry offers an attractive proteomic solution for analysis of membrane protein complexes and for basic expression profiling.