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CHANDRAN V RMass SpectrometryShotgun proteomicsNot availablePlanktonicLc-ms/msBiofilmEnterococcus faecalis sk460http://www.ebi.ac.uk/pride/archive/projects/PXD010770The strain Enterococcus faecalis SK460 is a strong biofilm former and was isolated from chronic diabetic foot infection. The strain was cultured in planktonic (12hr, 24hr) and biofilm stages (12hr, 24hr) for protein isolation to carry out quantitative proteome analysis. The cell pellet obtained from different planktonic and biofilm stages were resuspended in PBS (1ml PBS per gram of cells) and transferred to 2ml microcentrifuge tubes containing glass beads (0.5mm), Protease inhibitor (1mM PMSF) and incubated on ice for 5 minutes. The tubes were then placed in a Mini Bead beater (BioSpec Products, USA) and subjected to three, one minute pulses at 4200 rpm with 1 min interval. The suspensions were centrifuged for 10 min at 13,000 rpm at 4 °C and the supernatant was collected. Protein concentration was estimated by bicinchoninic acid protein assay kit (Thermo Scientific, USA). 100 micrograms of protein from each sample were used for in-solution trypsin digestion. DL-dithiothreitol (10 mM) (Sigma-Aldrich, USA) in 50mM ammonium bicarbonate (Sigma-Aldrich, USA) buffer at 60ᴼC for 30 minutes was used to reduce the disulfide bonds. The subsequent alkylation of proteins was done with 200mM iodoacetamide (Sigma-Aldrich, USA) in the same buffer at 27°C for 30 minutes in the dark. The alkylated proteins were digested with trypsin (sequencing-grade modified trypsin, Sigma-Aldrich, USA; 1:25 w/w) in 50mM ammonium bicarbonate buffer by incubating overnight at 37ᴼC. Formic acid(Sigma-Aldrich, USA; 1% v/v) was used to terminate the trypsin digestion reaction. The digested peptide solution was centrifuged at 14,000 rpm at 4ºC for 12 minutes, and the supernatant was stored at -20ᴼC until the LC/MS/MS analysis. The peptide samples were analyzed by using a nano ACQUITY UPLC® System (Waters, UK) coupled to a Quadrupole-Time of Flight (Q/TOF) mass spectrometer (SYNAPT-G2, Waters, UK) controlled by MassLynx4.1 SCN781 software (Waters, UK). In each sample, equivalent to 3µg of protein (3µl), was injected in ‘partial loop’ mode and was loaded into the reverse phase column with 0.1% formic acid in water as mobile phase A, and 0.1% formic acid in acetonitrile as mobile phase B, using the binary solvent manager. The trap column (Symmetry® 180µm x 20mm C18 5µm, Waters, UK) was used to remove any salt by employing a high flow rate (15µl/minute) with 99.9% mobile phase A and 0.1% mobile phase B for 1 minute. The peptide separation was performed in a 75µm x 100mm BEH C18 column (Waters, UK), with a particle size of 1.7µm. Gradient elution with 1-40% mobile phase B, for 55.5 minutes at 300nl/minute flow rate, was employed. After separation, the column was washed with 80 % mobile phase B for 7.5 minutes and re-equilibrated with 1% mobile phase B for 20 minutes. The column temperature was maintained at 40ᴼC. Three technical replicate runs were performed for each sample. Peptides eluted from the nano-LC were monitored by SYNAPT® G2 High Definition MS™ System (HDMSE System (Waters, UK). The time of flight analyzer (TOF) was calibrated with a solution of 500fmole/µl of human [Glu1]-Fibrinopeptide B (Sigma Aldrich, USA), and the lock mass acquisition was performed every 30 seconds by the same peptide delivered through the reference sprayer of the nano-LockSpray source at a flow rate of 500nl/minute. This calibration sets the analyzer to detect ions in the range of 50 - 2,000 m/z. The mass spectrometer was operated in the ‘resolution mode’ with a resolving power of 18,000 FWHM, and the data acquisition was performed in ‘continuum’ format. The data were acquired by rapidly alternating between two functions – Function-1 (low energy) and Function-2 (high energy-MSE). Each spectrum was acquired in 0.9 seconds with an interscan delay of 0.024s.PrideSpectrum countingPeptide countingCarbamidomethylOxidationThe acquired ion mobility enhanced MSE spectra was analysed using ProteinLynx Global SERVER™ v3.0.2 (PLGS, Waters) for protein identification as well as for the label-free relative protein quantification. Data processing includes lock mass correction post acquisition. Processing parameters for PLGS were set as follows: noise reduction thresholds for low energy scan ion – 150 counts, high energy scan ion - 50 counts and peptide intensity - 500 counts (as suggested by manufacturer). The protein identifications were obtained by searching against Enterococcus faecalis V583 (NCBI Reference Sequence: NC_004668.1). During database search, the protein false positive rate was set to 1%. The parameters for protein identification was made in such a way that a peptide was required to have at least 1 fragment ion match, a protein was required to have at least 3 fragment ion matches and a protein was required to have at least 1 peptide match for identification. Oxidation of methionine was selected as variable modification and cysteine carbamidomethylation was selected as a fixed modification. Trypsin was chosen as the enzyme used with a specificity of 1 missed cleavage. Data sets were normalized using the ‘auto-normalization' function of PLGS and label-free quantitative analyses was performed by comparing the normalized peak area/intensity of identified peptides between the samples. Furthermore, only a fold change higher than 50% difference (ratio of either <0.50 or >2) was considered to be indicative of significantly altered levels of expression.ProteomicsDr.Sabu ThomasSynapt MSscientist EII , Rajiv Gandhi centre for BiotechnologyCOMPLETEEnterococcus Faecalis (streptococcus Faecalis)maheshchandran@rgcb.res.in31253082 Suryaletha K, Narendrakumar L, John J, Radhakrishnan MP, George S, Thomas S. Decoding the proteomic changes involved in the biofilm formation of Enterococcus faecalis SK460 to elucidate potential biofilm determinants. BMC Microbiol. 2019 19(1):146 10.1186/s12866-019-1527-2BiologicalTechnician, Mass spectrometry and proteomics core facility, RGCBIndia10.6019/PXD010770<h4>Background</h4>Enterococcus faecalis is a major clinically relevant nosocomial bacterial pathogen frequently isolated from polymicrobial infections. The biofilm forming ability of E. faecalis attributes a key role in its virulence and drug resistance. Biofilm cells are phenotypically and metabolically different from their planktonic counterparts and many aspects involved in E. faecalis biofilm formation are yet to be elucidated. The strain E. faecalis SK460 used in the present study is esp (Enterococcal surface protein) and fsr (two-component signal transduction system) negative non-gelatinase producing strong biofilm former isolated from a chronic diabetic foot ulcer patient. We executed a label-free quantitative proteomic approach to elucidate the differential protein expression pattern at planktonic and biofilm stages of SK460 to come up with potential determinants associated with Enterococcal biofilm formation.<h4>Results</h4>The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of proteomic data revealed that biofilm cells expressed higher levels of proteins which are associated with glycolysis, amino acid biosynthesis, biosynthesis of secondary metabolites, microbial metabolism in diverse environments and stress response factors. Besides these basic survival pathways, LuxS-mediated quorum sensing, arginine metabolism, rhamnose biosynthesis, pheromone and adhesion associated proteins were found to be upregulated during the biofilm transit from planktonic stages. The selected subsets were validated by quantitative real-time PCR. In silico functional interaction analysis revealed that the genes involved in upregulated pathways pose a close molecular interaction thereby coordinating the regulatory network to thrive as a biofilm community.<h4>Conclusions</h4>The present study describes the first report of the quantitative proteome analysis of an esp and fsr negative non gelatinase producing E. faecalis. Proteome analysis evidenced enhanced expression of glycolytic pathways, stress response factors, LuxS quorum signaling system, rhamnopolysaccharide synthesis and pheromone associated proteins in biofilm phenotype. We also pointed out the relevance of LuxS quorum sensing and pheromone associated proteins in the biofilm development of E. faecalis which lacks the Fsr quorum signaling system. These validated biofilm determinants can act as potential inhibiting targets in Enterococcal infections.Decoding the proteomic changes involved in the biofilm formation of Enterococcus faecalis SK460 to elucidate potential biofilm determinants.Suryaletha Karthika K, Narendrakumar Lekshmi L, John Joby J, Radhakrishnan Megha Periyappilly MP, George Sanil S, Thomas Sabu Ssodium salt, BBE, ammonium formate, 13C-labeled, Size, IL1BC, cadmium salt, MeCN, ion, Streptococcus Group D, CASP-1, Particle, FEZL, Foot Ulcer, magnesium formate, monosomy 2q32, Apaf-1, zinc salt, Solvent, Long Term, Polypeptides, protein polypeptide chains, KL receptor activity, M(3L)i, cobalt(II) formate dihydrate, CH3-C#N, ionic compounds, B1, SCO5, 1, 2, Msal-1, SCO1, Software Engineering, Il1bc, Gsfsow3, WMS, ASD2, (R*, strong, sarcoma of breast, hac-1, M, weight-weight percentage, Salt, Msal, proteins, W, Salz, sel, ecotype, D17Mit170, IL-1BC, salts, AI047805, isolation and purification, salt, g, column, sample, D1, n, Bs, monosomy 2q32-q33, Infections and Infestations, D9, M(3)RpS17, nickel salt, SGS, LC-MS2, Caspase-1 subunit p10, BZRP, cobalt (+2) salt, anon-53Fa, Longterm Effect, l(2)SH0173, breast sarcoma, Tl3, Tl2, not genetically inherited, Streptococcus glycerinaceus, Software Tools, PBS, DmelCG7788, ACMICD, PBR, Fibrinopeptides B, PBT, Computer Applications, common salt, sodium (4:1:1) salt, Dm1, magnesium salt, Streptococcus liquefaciens, increased period, M(3)i(55), SATB2-associated syndrome, Computer Applications Software, Foot, Del(2)(q32), Sizes, CG6829, Hydrogen Oxide, dark/hac-1/dapaf-1, rpS17, pbt, 2q32q33 microdeletion syndrome, Software Applications, ionic compound, Skeletal muscle enolase, Source Software, formate, Infection and Infestation, cromium (+3), beta Trypsin, human, w/w, Speed, Fibrinogen beta chain, Applications, Painful bladder syndrome, liquid chromatography-tandem mass spectroscopy, Drice, ammonium (4:1) salt, GSD13, methyl cyanide, DRICE, lead (+2) salt, krk1, Computer Software Applications, BPBS, Cleland Reagent, Particle Sizes, nickel formate dihydrate, not genetically inherited., lead formate, sarcoma of the breast, Effects, DrIce, DrICE, aluminum salt, number, 4-dimercapto-, R*)-, protein-containing complex, thomson, Chalk, body system, CG7826, mass-to-charge ratio, period, SATB2 syndrome, M(3)i, isolation, TOF, tough, LC-MSMS, CG7835, Gene Products, technical_replicate, CG42273, system, ZNF312, Application, Open Source Softwares, anatomical systems, thallium (+1) salt, Longterm, Software Application, PCE-2, beta-Trypsin, rubidium salt, Open Source Software, pk18, Long-Term, l(3)dtOA4, methanoic acid, ions, man, MBR, Solution, Computer Software Application, Tools, Caspase-1 subunit p20, Kochsalz, M(3)i[55], PTBR, Chronic, peptidos, PTHB1, proto-oncogene c-Kit, dark/dapaf-1/hac-1, mass percentage, cou, 1-(14)C-labeled, caspase 3, dApaf-1, Proteins, 2-phospho-D-glycerate hydro-lyase, Clelands Reagent, potassium formate, function, CE-2, copper (+2) salt, increased time, acetonitrile, buffer, chronic, Cell, Tool, polypeptide, APAF1, Lr, native protein, Temperatures, Natriumchlorid, Hac-1, KIT ligand receptor activity, C18, cupric formate, chemical analysis, Long Term Effects, high time, BcDNA:RE44119, MFS1, FWHM, NaCl, aluminum formate, XKrk1, Cleland's, AU020952, technical replicate, lithium formate, M(3)q, Dark/Dapaf-1/HAC1, B130022O04Rik, IL-1 beta-converting enzyme, ethanenitrile, Interleukin-1 beta-converting enzyme, Muscle-specific enolase, CD117, mDRC, anon-EST:Posey48, Enterocoque, Longterm Effects, ME-IV, Cleland's Reagent, Computer Programs, Gene Proteins, ZFP312, Micrococcus ovalis, Applications Softwares, C-Kit, Dietary Sodium, Ssm, Feet, M(3L)i[55], xkl-1, Bra, liquid chromatography tandem mass spectrometry, quantitative, liquid chromatography tandem mass spectroscopy, Hac-1/Dark, Rps17, Meth, NCMe, determination, DmelCG6829, RPS17, zinc formate, Infestations and Infections, CG7788, crice, lead salt, protein, Xkl-1, Alkylations, M(3)67, peptide, Gsfsco1, ammonium tetraformate, halite, ClvPrd, peptido, Min, Apaf1, protein aggregate, monosomy 2q32q33, Gsfsco5, ARK, Effect, Computer Program, DmelCG42273, SOW3, me75, LCMSMS, peptides, reference sample, TACHD, l(3)67BDo, Open, min, arc, Computer Programs and Programming, mAPC, purification, IBP, analyzer, copper, Estimated, ark, T1, cloruro sodico, reaction, chromosome 2q32-q33 deletion syndrome, lithium salt, Sputolysin, Ice, ICE, dApaf1, Bladder pain syndrome, Sl, GPHYSD2, ammonium (2:1) salt, Long-Term Effects, rock salt, PLATEST, ice, iCE, D-Apaf-1, drice, LC-MS/MS, M(3)67C, Dmel_CG7826, Tr-kit, Th, Thermo Scientific, Source Softwares, Programs, glass, Program, Cleland, Ionen, drIce, drICE, Computer Applications Softwares, kl1-A, cultivar, Softwares, KIT, Biofilm, nickel (+2) salt, Dmel_CG7835, Crystal, Mnb, MNB, tyrosine-protein kinase Kit, ammonium salt, Painful Bladder Syndrome, Spalt, P45, Rp S17, cobaltous formate, Del(2)(q32q33), 3-Butanediol, kit, hac1, MASS, AW124434, natrii chloridum, organ system, dapaf-1S, apaf-1, dapaf-1L, Acetamide, copper salt, p45, Polypeptide, Fibrinopeptide B, Proteomes, time, Platelets, Bladder Pain Syndrome, Diabetic Feet, human being, Dapaf-1/HAC-1, Reagent, cesium salt, SCF receptor activity, FBN, Eno-3, Gene, Enterococcus proteiformis, slow time, Computer, presence, LC-MS-MS, Buffer, formic acid, Dark/Hac-1/dApaf1, Hac1, M(3)S33, potassium salt, Dark/Hac-1/dApaf-1, resilient, polypeptide chain, scfr, glass syndrome, ECTOL1, 14C-labeled, Tina, Low, Disulfide, SCFR, Pulses, 4.2.1.11, Fdc, Glass, iones, dapaf-1, chlorure de sodium, Minute, dapaf, dark, calcium formate, MSE, OCTD, Infestation and Infection, DmelCG3922, Suspension, DARK, sels, cromium (+3) salt, Long-Term Effect, dArk, Dapaf-1, FEZ, 2-iodo-, sodium formate, Controlled, Applications Software, 2q32-q33 microdeletion syndrome, Open Source, DYRK1, Controlling, data, bead, Computer Software, nickel formate, DBI, 2q32q33 microdeletion syndromes, 3H-labeled, protein complex, sales, total expressed protein, strontium formate, apaf1, strontium salt, Peptide, strain, count in organism, LC/MS/MS, Software Tool, natural protein, PKBS, Protein, Streptococcus faecalis, Infection, table salt, Dyrk1, prolonged period, CES2A1, Dark, connected anatomical system, Micrococcus zymogenes, Software, Dark/Apaf-I, ACETONITRILE, cyanomethane, WMS2, Sal, VSD1, SAS, c-KIT, 10^[-9], Diabetic Foot Ulcer, CC1, Engineering, Tripcellim, Salze, Diabetic, high flow, dApaf-1/DARK/HAC-1, sample population, chromic formate, CG3922, sal, Interleukin-1 beta convertase, Protein Gene Products, Trypure, c-kit, Ion, Liver regeneration-related protein LRRG036|LRRG043|LRRG189, SSKS, 3.4.22.36, Calibrations, calcium salt, Peptid, assay, dAPAF-1, Enolase 3, presence or absence in organism, sodium chlorideMicrococcus ovalis, Enterococcus proteiformis, Biofilm., Micrococcus zymogenes, Streptococcus liquefaciens, Streptococcus glycerinaceus, Enterocoque, Streptococcus Group D, Streptococcus faecalisBBE, IPP2A2, 2-amino-4-(methylsulfanyl)butanoic acid, Cysteine Hydrochloride, ion, False, Ass-1, number, L Isomer, cleavage, Eno-3, FBN, protein, protein-containing complex, presence, PHAPII, 5730420M11Rik, peptide, Polypeptides, protein polypeptide chains, L-Isomer Methionine, Methionine, ClvPrd, peptido, polypeptide chain, template-activating factor I, ECTOL1, AA408052, Plgs, L-Methionine, fold, Low, Half-Cystine, protein aggregate, WMS, Pedameth, 4.2.1.11, SET, proportion, me75, F, peptides, TAF-I, iones, M, L Cysteine, phosphatase 2A inhibitor I2PP2A, ipp2a2, beta-Trypsin, 2pp2a, proteins, ions, SCAN, free, D17Mit170, CG10574, T1, MSE, OCTD, DmelCG4299, ASS, set, IGAAD, Enterococcus faecalis str. V583, data processing, 2PP2A, 2-amino-4-(methylthio)butanoic acid, DmelCG10574, taf-ibeta, dSET, dSet, peptidos, Half Cystine, GPHYSD2, metionina, Racemethionine, SGS, ratio, phapii, data, Noise, cou, DL-Methionine, Zinc Cysteinate, AI447866, protein complex, igaad, 2-Amino-4-(methylthio)butyric acid, 2-phospho-D-glycerate hydro-lyase, Pollution, StF-IT-1, ratio., function, Tl3, L-Isomer, Tl2, not genetically inherited, Peptide, ACMICD, polypeptide, count in organism, Lr, native protein, Ionen, natural protein, I-2PP2A, Protein, Dm I-2, I2PP2A, sequence, MFS1, Methionin, methionine, Hmet, WMS2, Data Base, Met, HLA-DR-associated protein II, DI-2, Skeletal muscle enolase, I-2Dm, proportionality, Tripcellim, rate, L-Cysteine, MASS, CG4299, Muscle-specific enolase, inhibitor of granzyme A-activated DNase, beta Trypsin, primary structure of sequence macromolecule, Noises, I-2PP1, Trypure, data analysis, dSET/TAF-Ibeta, 2610030F17Rik, 1110030K05Rik, TAF-IBETA, Ion, SSKS, Bra, quotient, alpha-amino-gamma-methylmercaptobutyric acid, Peptid, TAF-Ibeta, GSD13, Noise Pollution, Polypeptide, quantitative, AA407739, i2pp2a, Liquimeth, Enolase 3, presence or absence in organismForms, study, viral infection, Infections, protein complex, Wound, Streptococcus Group D, number, Infestations and Infections, Enterococcus proteiformis, Infection and Infestation, virus process, present in organism, proteins, protein, protein-containing complex, infectivity, presence, Streptococcus glycerinaceus, Enterocoque, Infestation and Infection, count in organism, protein polypeptide chains, Micrococcus ovalis, native protein, Wound Infections, natural protein, polypeptide chain, Streptococcus liquefaciens, Protein, Streptococcus faecalis, Infection, Pathogenicity, virulence, Infections and Infestations, quantitative, Biofilm., associated, Micrococcus zymogenes, Biofilm, protein aggregate, presence or absence in organismbiochemical pathways, Signal Transductions, Metabolic Process, Gene Ontology Projects, Materials, cell-cell signaling involved in quorum sensing, single-organism developmental process, determination, 2410041A17Rik, Allelochemical, ACTG, ACTE, Streptococcus Group D, Metabolic Concepts, Infestations and Infections, Embden-Meyerhof, protein, fs(1)M34, Embden Meyerhof Parnas Pathway, infectivity, homolog of separase, Act5, csp2, Long Term, l(1)G0420, DmelCG12051, Receptor Mediated Signal Transduction, protein polypeptide chains, primary metabolites, 5-dihydroxypentan-2, Roles, Quorum Quenchings, F15I1.12, responsivity, Quenching, Concepts, Project, HEL-176, CG4601, Embden-Meyerhof Pathway, TASTY, Metabolism Concept, Cell Signaling, protein aggregate, Phenomenon, Effect, drug resistance, IKKg, KEY, Key, F15I1_12, multicellular organismal biosynthetic process, signal transduction by protein phosphorylation, A, strong, viral infection, C, single-organism biosynthetic process, cyt5C, Genomes, DHO, ERV-R envelope protein, catabolism, anabolism, CG18572, Pathways, virus process, present in organism, proteins, metabolic process resulting in cell growth, Ectohormones, ecotype, free, cellular amino acid biosynthetic process, amino acid anabolism, LuxS, SU, RADIALLY SWOLLEN 4, Gelatinase, act 42A, signaling process, Role Concepts, Ac5C, CG4027, biotransformation, ACT, Act, Signal Transduction Pathways, Infections and Infestations, Transductions, associated, TM, Catabolism, Synomones, Long-Term Effects, Envelope polyprotein, HERV-T Env protein, single organism signaling, Pathway, l(1)G0330, Process, Actin/BAP47, CTE-II, metabolism resulting in cell growth, AACT, signal transduction by conformational transition, anaerobic glycolysis, Longterm Effect, CTE-IIa, act, Ach1, hBACH, ACH1, RSW4, Embden-Meyerhof-Parnas, LACH1, Streptococcus glycerinaceus, HERV-T_19q13.11 provirus ancestral Env polyprotein, results, signal transduction by trans-phosphorylation, act42A, Transmembrane protein, Embden-Meyerhof-Parnas Pathway, GIG25, DmIKKgamma, signaling cascade, Role Concept, Pheromone, Streptococcus liquefaciens, increased period, Alpha-1-antichymotrypsin His-Pro-less, Su(b), dIKK, AFFX-Dros-ACTIN_M_r_at, Kenny, Role, Pathogenicity, DFNA26, Genetic Materials, actin, Ontology Projects, secretion, cultivar, ribosylhomocysteinase activity, Biofilm, LACH, Transduction, DFNA20, GAT, Genetic Material, PCR, Semiochemicals, Quorum Quenching, Quenchings, Gene Ontologies, Ontology, DmelCG4027, Ontologies, Projects, System, l(1)G0117, IKK-gamma, Infection and Infestation, Actin, ERV3 envelope protein, ERV-3 envelope protein, beta-actin/Bap47, Embden-Meyerhof pathway, organ system, l(1)G0486, Phenotypes, l(1)G0245, HERV-R_7q21.2 provirus ancestral Env polyprotein, actin5C, DmelCG16910, glycolysis, signaling pathway, l(1)G0009, Material, ACTL3, Lach1, metabolites, rhamnose formation, Cistron, BACH, Embden-Meyerhof Pathways, time, Proteomes, rhamnose anabolism, l(1)Ab, Act5c, Signal Transduction Systems, Diabetic Feet, biological signaling, l(1)G0010, bioformation, Effects, Processes, Allomone, drug susceptibility/resistance, Receptor-Mediated, number, secondary metabolites, ACT5C, Gene, CG12051, Enterococcus proteiformis, Signal Transduction System, slow time, biosynthesis, rhamnose synthesis, Serpin A3, Metabolic Processes, protein-containing complex, Bach, body system, presence, dIKK-gamma, Gene Ontology Project, Gene Ontology, Quorum, period, l(1)G0025, Signal Transduction, resilient, polypeptide chain, Metabolism, DmIKK-gamma, tough, amino acid biosynthesis, Gene Products, system, Cell growth-inhibiting gene 24|25 protein, dmIKKgamma, IKK[[gamma]], epithiospecifier protein, Metabolism Phenomena, study, ERV3-1 envelope protein, reactivity, act5C, anatomical systems, pattern, Genetic, SEPARASE, formation, Longterm, distribution, Allelochemicals, BRWS2, Signal, CG7005, Metabolic Concept, Signal Pathways, Semiochemical, Long-Term, Infections and Infestations., arginine metabolism, Signal Transduction Pathway, amino acid formation, synthesis, fs(1)829, anon-EST:fe2D2, Act42a, Infestation and Infection, signalling pathway, detection of cell density by secreted molecule, ESR, 1700027G07Rik, IKK, time of survival, DmelCG18572, HERV-R envelope protein, signal transduction by cis-phosphorylation, Chronic, Allomones, Long-Term Effect, T11, 3-dione-forming], Kairomones, Signal Pathway, Act-5C, amino acid synthesis, modifed Embden-Meyerhof pathway, Cte-II, S-ribosylhomocysteinase activity, l(1)G0177, quorum sensing system, 42A, data, Encyclopedias, CPS, Embden Meyerhof Pathway, Sensing, degradation, Drug resistance, protein complex, synthesize, Proteins, dJ393D12.2, F7H19.150, Receptor-Mediated Signal Transductions, total expressed protein, 3.2.1.148, Surface protein, increased time, Cistrons, Cell, chronic, AESP, Concept, strain, Metabolic Phenomena, development, IKKgamma, Metabolism Concepts, count in organism, survival, CAD, F7H19_150, native protein, natural protein, beta-actin, Receptor-Mediated Signal Transduction, Kairomone, Systems, Protein, Long Term Effects, chemical analysis, Streptococcus faecalis, Phenomena, M32055, Infection, Resistance, high time, Act42, background, prolonged period, connected anatomical system, ACTA3, Micrococcus zymogenes, signalling cascade, metabolism, Metabolic Phenomenon, Ontology Project, l(1)G0079, multicellular organism metabolic process, rhamnose biosynthesis, biodegradation, EPITHIOSPECIFYING SENESCENCE REGULATOR, Metabolic, Dmikkgamma, Diabetic Foot Ulcer, PYR1, VSCM, response to drug, act 5C, metabolite, patient, S-(5-deoxy-D-ribos-5-yl)-L-homocysteine homocysteine-lyase [(4S)-4, CG16910, Embden-Meyerhof-Parnas pathway, Enterocoque, introduction, Longterm Effects, signalling, Drug, Protein Gene Products, DmelCG7005, Gene Proteins, Micrococcus ovalis, single-organism metabolic process, signalling process, DRORUD, virulence, BAP47, assay, ACTSG, quantitative, response, Bap47, Actin5C, presence or absence in organism, Anabolismcount in organism, Micrococcus ovalis, determination, Streptococcus liquefaciens, chemical analysis, Streptococcus Group D, Streptococcus faecalis, number, total expressed protein, assay, Enterococcus proteiformis, quantitative, Biofilm., Micrococcus zymogenes, Proteomes, presence, Streptococcus glycerinaceus, Enterocoque, presence or absence in organismfalseQuantitative proteome analysis of Enterococcus faecalis SK460 during planktonic and biofilm stagesEnterococcus faecalis is a major nosocomial pathogen frequently isolated from non- healing wound infections. The major factor responsible for the virulence and establishment of infection is its ability to form robust biofilm. This renders the recalcitrant nature of Enterococci towards the current treatment strategies. In the present study, the quantitative proteomic approach is carried out to elucidate the protein expression levels in E. faecalis at planktonic and biofilm stages. This will help to identify biofilm associated pathways in E. faecalis which inturn can be considered as novel targets for biofilm 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