Thus, H. influenzae must adapt to the growth GANT61 and metabolic conditions in multiple microenvironments and bacterial cells may express different proteins, depending on the microenvironment.Proteomic profiling of sputum-grown cells may represent an average of multiple conditions. Table 1 Proteins of the glycolysis pathway identified in H. influenzae strain 11P6H identified during growth in pooled human sputum Protein ID# Identified Protein Gene Genome ID numbera Molecular Weight CDMb Sputumc 1237 phosphoenolpyruvate-protein
phosphotransferase fruA HI0446 64 kDa 100% (9.7%) 100% (11%) 412 Fructose specific phosphotransferase system IIA/HPr components fruB HI0448 53 kDa 100% (24%) 100% (8.2%) 1149 Aldose 1-epimerase galM HI0818 38 kDa 100% (11%) 100% (15%) 423 1-phosphofructokinase fruK HI0447 34 kDa 100% (24%) 100% (15%) 557 6-phosphofructokinase pfkA HI0982 23 kDa 100% (21%) 95% (20%) 57 Fructose-bisphosphate aldolase fba HI0524 39 kDa 100% (47%) 100% (36%) 657 glucose-6-phosphate isomerase pgi HI1576 37 kDa 100% (19%) 100% (15%) 392 Triosephosphate isomerase tplA HI0678 27 kDa 100% (25%) 100% (19%) 97 Glyceraldehyde-3-phosphate mTOR activity dehydrogenase gapA HI0001 36 kDa 100% (40%) 100% (39%) 111 3-phosphoglycerate
kinase pgk HI0525 41 kDa 100% (39%) 100% (37%) 34 phosphoglyceromutase gpmA HI0757 26 kDa 100% (52%) 100% (56%) 79 enolase eno HI0932 46 kDa 100% (43%) 100% (32%) 133 Pyruvate kinase pykA AZD5153 order HI1573 49 kDa 100% (37%) 100% (47%) 538 Dihydrolipoamide acetyltransferase aceF HI1232 57 kDa 100% (22%) 100%
(23%) 305 Pyruvate dehydrogenase subunit E1 aceE HI1233 99 kDa 100% (28%) 100% (30%) aID numbers based on annotation of H. influenzae strain KW20 Rd http://cmr.jcvi.org/cgi-bin/CMR/GenomePage.cgi?org=ghi bProtein probabilities values as calculated by Proteinprophet algorithm for proteins detected during growth in chemically define media (CDM). Number in parentheses represents the sequence coverage expressed by the percentage (-)-p-Bromotetramisole Oxalate of amino acid residues identified. All peptides were filtered with a set of criteria as specified in the Methods. CDM cProtein probabilities for proteins detected during growth in 20% pooled human sputum. Proteins expressed in increased amount during growth in sputum Additional File 3 lists the 31 proteins that were present in a ratio of > 1.5 in sputum-grown compared to media-grown bacteria, along with the corresponding gene and the COG functional category.A range of proteins is represented but clear-cut themes are observed and these are shown graphically in Figure 2 and are outlined below. Figure 2 Distribution of functional categories of 31 proteins that were present in increased abundance during growth of H. influenzae in 20% pooled human sputum compared to growth in chemically defined media.