Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/proteinGroups.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/experimentalDesignTemplate.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_17.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_13.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_04.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_22.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_25.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_08.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_16.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_03.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_10.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_23.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_07.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_15.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_15.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_06.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_11.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_11.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_20.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_06.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_14.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_14.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_20.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_12.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_12.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_05.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_05.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_21.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_13.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_21.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_04.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_17.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_22.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_08.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_25.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_16.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_03.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_23.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_24.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_10.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_07.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_24.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_19.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_19.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_18.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_18.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_09.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_09.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_1_26.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/JG_QL109_2_26.RAWftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/ql109_all_orfs.fastaftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/10/PXD006577/maxquant_txt_results.zipprimaryOK200101700g.wezel@biology.leidenuniv.nlJacob GubbensMass SpectrometryShotgun proteomicsNot availableSiderophoreLc-msmsNatural productsStreptomycesDimethyl labelinghttp://www.ebi.ac.uk/pride/archive/projects/PXD006577The Streptomyces sp. genome sequence has been published, GenBank accession number: LNBE00000000. Here, Streptomyces sp. MBT76 was grown in liquid NMMP medium containing 1% (w/v) glycerol and 0.5% (w/v) mannitol as carbon sources, but lacking polyethylene glycol. This basic NMMP medium were perturbed by using four different additives (or no additive) to create varying growth conditions: (A) no additive, (B) 2% (w/v) NaCl, (C) 1% (w/v) starch, (D) 0.8% (w/v) Bacto peptone (Difco), (E) 0.6% (w/v) Bacto yeast extract (Difco). All the cultures of Streptomyces sp. MBT76 were incubated at 30 °C for 72 hours, with constant shaking at 220 rpm. Mycelium was washed in 2 ml of 100 mM Tris/HCl buffer (pH 7.5) supplemented with 10 mM MgCl2, and then resuspended in 5 ml of ice cold acetone for 10 min. After removal of acetone by centrifugation, the pellets were resuspended in 1 ml of 1% (w/v) SDS for 2 min. The resulting protein suspension was separated from the debris by centrifugation, and the protein concentrations were determined using the Qubit fluorometric system (Thermo) with BSA standard. Some 167 µg of protein was precipitated for each sample using chloroform/methanol and then dissolved using RapiGest SF surfactant (Waters, Milford MA). The proteins were further digested with trypsin after iodoacetamide treatment, and the resulting primary amines of the peptides were dimethyl labeled using three combinations of isotopomers of formaldehyde and cyanoborohydride on Sep-Pak C18 200 mg columns (Waters, Milford MA), via CH2O + NaBH3CN (light, condition A (exp. 1), condition C (exp. 2)), CD2O + NaBH3CN (medium, condition D (exp. 1 & 2)), and 13CD2O + NaBD3CN (heavy, condition B (exp. 1), condition E (exp. 2). Light-, medium-, and heavy-labeled peptides with 4 Da mass differences were mixed 1:1:1 to obtain 0.5 mg for fractionation by cationic exchange (SCX) chromatography using a polysulfoethyl A column (PolyLC, 100 x 2.1 mm, particle size 5 m, average pore size 200 Å). Mobile phases for SCX chromatography consisted of solvent A (10 mM KH2PO4, 20% (v/v) acetonitrile, pH 3) and solvent B (10 mM KH2PO4, 20% (v/v) acetonitrile, 0.5 M KCl, pH 3). The running program for SCX was a gradient of 0-18% solvent B in 18 CV (column volume), 18-30% solvent B in 6 CV, and 30-100% solvent B in 5 CV, at a constant flow rate of 250 μl/min. In total, 24 peptide fractions were collected. Acidified peptides were desalted using StageTips. To remove acetonitrile, samples were concentrated in a vacuum concentrator. Prior to LC-MS analysis, the volume was adjusted to 70 μl using a 3% (v/v) acetonitrile, 0.1% (v/v) formic acid solution. Peptides were analyzed on a Surveyor nanoLC system (Thermo, Waltham, MA) hyphenated to an LTQ-Orbitrap mass spectrometer (Thermo). Gold and carbon coated emitters (OD/ID = 360/25 μm tip ID = 5 μm), trap column (OD/ID = 360/100 μm packed with 25 mm robust Poros10R2/15 mm BioSphere C18 5 μm 120 Å) and analytical columns (OD/ID = 360/75 μm packed with 20 cm BioSphere C18 5 μm 120 Å) were obtained from Nanoseparations (Nieuwkoop, the Netherlands). The mobile phases [A: 0.1% (v/v) formic acid/H2O, B: 0.1% (v/v) formic acid/acetonitrile] were made with ULC/MS grade solvents (Biosolve, Valkenswaard, the Netherlands). The emitter tip was coupled end-to-end with the analytical column via a 15 mm long TFE Teflon tubing sleeve (OD/ID 0.3 × 1.58 mm, Supelco, St Louis, MO) and installed in a stainless steel holder mounted in a nano-source base (Idex, Northbrook, IL). General mass spectrometric conditions were as follows: an electrospray voltage of 1.6–2.0 kV was applied to the emitter, no sheath and auxiliary gas flow, ion transfer tube temperature 150°C, capillary voltage 20 V, tube lens voltage 110 V. For shotgun proteomics analysis, 10 μl of the samples was pressure loaded on the trap column with a 10 μl min−1 flow for 5 min followed by peptide separation with a gradient of 95 min 7–25% B, at a flow of 300 μl min−1 split to 250 nl min−1 by the LTQ divert valve. For each data-dependent cycle, one full MS scan (300–2000 m/z) acquired at high mass resolution (60 000 at 400 m/z, AGC target 1 × 106, maximum injection time 1000 ms) in the orbitrap was followed by five MS/MS fragmentations in the LTQ linear ion trap (AGC target 1 × 104, maximum injection time 50 ms for mixture 1 of the glucose vs mannitol measurement, 120 ms for the other mixtures) from the five most abundant ions. MS2 settings were as follows: collision gas pressure 1.3 mT, normalized collision energy 35%, ion selection threshold of 750 counts, activation q = 0.25 and activation time of 30 ms. Fragmented precursor ions were dynamically excluded for 150 s and ions with z < 2 or unassigned were not analyzed. Exclusion width was set to −10 ppm to +10 ppm.PrideNot availableiodoacetamide derivatized residuemonohydroxylated residueData analysis was performed using MaxQuant 1.4.1.2 in one run. Variable modification was set to oxidation of methionine, fixed modification was set to carbamidomethylation of cysteine, and dimethyl labelling was selected as quantification method. MS/ MS spectra were searched against a database of translated coding sequences obtained from genome of Streptomyces sp. MBT76 with a false discovery rate of 1% for both proteins and peptides, and an additional second peptide search was performed. Quantification was performed according to MaxQuant standard settings. In short, a median protein expression ratio was determined using normalized expression ratios of positively identified peptides and of low-scoring versions of peptides already positively identified in a separate MS/MS event. No median ratio was determined when less than three peptide ratios were available.ProteomicsProf.dr. G.P. van WezelLTQ OrbitrapMolecular Biotechnology, Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE, The NetherlandsPARTIALStreptomyces Sp. Mbt76j.gubbens@biology.leidenuniv.nl28945067 Gubbens J, Wu C, Zhu H, Filippov DV, Florea BI, Rigali S, Overkleeft HS, van Wezel GP. Intertwined Precursor Supply during Biosynthesis of the Catecholate-Hydroxamate Siderophores Qinichelins in Streptomyces sp. MBT76. ACS Chem Biol. 2017 12(11):2756-2766 10.1021/acschembio.7b00597Leiden UniversityThe explosive increase in genome sequencing and the advances in bioinformatic tools have revolutionized the rationale for natural product discovery from actinomycetes. In particular, this has revealed that actinomycete genomes contain numerous orphan gene clusters that have the potential to specify many yet unknown bioactive specialized metabolites, representing a huge unexploited pool of chemical diversity. Here, we describe the discovery of a novel group of catecholate-hydroxamate siderophores termed qinichelins (2-5) from Streptomyces sp. MBT76. Correlation between the metabolite levels and the protein expression profiles identified the biosynthetic gene cluster (named qch) most likely responsible for qinichelin biosynthesis. The structure of the molecules was elucidated by bioinformatics, mass spectrometry, and NMR. The genome of Streptomyces sp. MBT76 contains three gene clusters for the production of catecholate-peptide siderophores, including a separate cluster for the production of a shared catecholate precursor. In addition, an operon in the qch cluster was identified for the production of the ornithine precursor for qinichelins, independent of primary metabolism. This biosynthetic complexity provides new insights into the challenges scientists face when applying synthetic biology approaches for natural product discovery.Intertwined Precursor Supply during Biosynthesis of the Catecholate-Hydroxamate Siderophores Qinichelins in Streptomyces sp. MBT76.Gubbens Jacob J, Wu Changsheng C, Zhu Hua H, Filippov Dmitri V DV, Florea Bogdan I BI, Rigali Sébastien S, Overkleeft Herman S HS, van Wezel Gilles P GPsodium salt, Water, Formol, ammonium formate, 13C-labeled, mannite, Bhlha41, SCX, ion, E-421, Glukose, CASP-1, Particle, SDH, Visible Light, magnesium formate, CycEI, zinc salt, Steels, Solvent, Osmofundin, WATER, HOH, SDS, Magnesium chloride, T22F8_160, Carbon-12, ATGPR7, PH 3, B1, 1, HCL-C, 2, 3, 4, 5, germacrene A synthase activity, sylvite, WMS, nup154, C, SET, Oelsuess, pore, i2pp2a., phosphatase 2A inhibitor I2PP2A, Oxomethane, Basen, number of, ethylene glycol homopolymer, DmelCG4299, glucose, salt, Stainless Steels, D1, n, Homo sapiens disease, F2C=CF2, SGS, nickel salt, Glyceritol, chronic obstructive lung disease [Ambiguous], polyoxyethylene, Caspase-1 subunit p10, cobalt (+2) salt, mannitol, Oxide, Longterm Effect, Light, Normalcy, tip, not genetically inherited, Expanded, DmelCG7788, ACMICD, CHRONIC OBSTRUCTIVE AIRWAY DIS, MgCl2, common salt, sodium (4:1:1) salt, tetrafluoro-, LIGHT, magnesium salt, Base2, Base1, (alpha-D)-Isomer, l(2)05206, activation, D-Glucose, E 421, Chronic obstructive lung disease, perfluoroethene, Gro, divided, DI-2, I-2Dm, formate, inhibitor of granzyme A-activated DNase, valva (Spanish, I-2PP1, light quantum, OBSTRUCTIVE PULMONARY DISEASE (COPD), duct, 5730422P05Rik, ammonium (4:1) salt, Monopotassium chloride, tlp, breadth, Oxides, Peptidomics, DrICE, number, 2-trans, CH3OH, CHRONIC OBSTRUCTIVE PULM DIS, Antp P1, precursor, Chronic obstructive pulmonary disease NOS, Antp P2, body system, SCXB, CG7826, SCXA, agua, Chronic obstructive pulmonary disease finding, C2F4, period, glycyl alcohol, PULMONARY DISEASE (COPD), Hu, CG7835, Gene Products, Cdi7, system, anatomical systems, Hydrogen chloride, thallium (+1) salt, Longterm, Lichtquant, Glycerin, PCE-2, rubidium salt, homopolymer, Fluoroplast, 4733401P19Rik, 2pp2a, ions, CG1028, CG10574, Sep-Pak C18, glycerolum, hairy cell leukemia, Formalin, Caspase-1 subunit p20, ATGRP7, ATGRP8, dSet, Chronic, peptidos, PTHB1, Magnesiumchlorid, 1-(14)C-labeled, chronic obstructive airways disease NOS, disorders, Methyl alcohol, copper (+2) salt, COLD - Chronic obstructive lung disease, DmAntp, carbone, buffer, CD258, ME2, (+)-(10R)-germacrene A synthase activity, CH2O, carbono, Kaliumchlorid, formaldehyde, l(2)k02514, Temperatures, native protein, DmCycE, C18, cupric formate, chemical analysis, Long Term Effects, condition, MFS1, Sodium, chronic airway obstruction, NaCl, chronic obstructive, aluminum formate, Perfluoraethylen, dJ142L7.2, AU020952, torn, Ant, GORE TEX, HVEM-L, GR-RBP7, GR-RBP8, CG4579, SWDS, ethanenitrile, Interleukin-1 beta-converting enzyme, polyethylene oxide, Polyethylene Oxide, Ns, PTFE, Photoradiations, Oxomethylene, chronic obstructive lung disease (disorder), quantitative, bHLHb19, Methoxide, channel-forming toxin activity, 2-tetrafluoroethylene, T22F8.160, NCMe, selection process, Chronic Airflow Obstructions, polyethylene, Antp1, classic hairy cell leukaemia, zinc formate, CG7788, Amylum, crice, KCl, lead salt, protein, clefted, ethylene glycol polymer, peptide, ammonium tetraformate, Glycerol, peptido, ClvPrd, Klotrix, Chronic Obstructive Pulmonary Disease (COPD), MBNL, protein aggregate, Injectables, wood alcohol, human disease, COLD (chronic obstructive lung disease), peptides, NEC in ICD9CM_2006, TAF-I, 5R)-Hexane-1, E430039A18Rik, mAPC, Myceliums, manna sugar, DL-glucose, Chronic airflow limitation, omega-hydroxypoly(ethylene oxide), BG:DS07700.1, anatomical valve, InChI=1/CH2O/c1-2/h1H2, IGAAD, lithium salt, DmelCG10574, ICE, GLYCINE RICH PROTEIN 7, discontiguous, C16orf53, blood capillary, dihydrogen oxide, GPHYSD2, ammonium (2:1) salt, Long-Term Effects, CHRONIC, (+)-germacrene A synthase activity, Tube, obstructive lung disease, Alcohol, iCE, drice, Nucleobase, ZC2HC5, [MgCl2], Dmel_CG7826, AI836084, StF-IT-1, HTATIP1, Osmitrol, fragmented, Airflow Obstructions, Aus, 6-hexol, drIce, CHRONIC OBSTRUCTIVE, BB114693, glycerol, tetrafluoroethylene, Polyaethylenglykol, Kaon-Cl 10, Goretex, GAS, nickel (+2) salt, Dmel_CG7835, CHRONIC OBSTRUCTIVE LUNG DIS, MNB, Yeast, ammonium salt, Tetrafluoraethen, EXP40, MS/MS, EXP42, cracked, P45, cobaltous formate, CG4299, AW124434, Stainless, surfactant, acqua, Trichloromethane, disease, Vitreous Carbon, Wood, EXP35, COPD - Chronic obstructive pulmonary disease, copper salt, Base, p45, Wasser, time, Carbowax, Amylomaize Starch, cesium salt, carbon, cycE, Cornstarch, Methylalkohol, PHAPII, CYCLE, Cold, Polyethyleneoxides, Propanetriol, polypeptide chain, template-activating factor I, ECTOL1, Injection, 14C-labeled, Wood Alcohol, light, ITF2, ANTC, Cold Temperatures, antp, l(2)10432, CYCE, ITF-2, 6-hexaol, DmelCG3938, MS2, Formaldehyde, CyclE, D-Mannitol, ipp2a2, Polyoxyethylene, 3938, calcium formate, l(2)k05007, dm-cycE, Suspension, wood naphtha, has or lacks parts of type, disorder, Tarflen, Long-Term Effect, l(2)01501, ANTP, polysulfoethylaspartamide, 2-iodo-, HHT1, COAD - Chronic obstructive airways disease, DYRK1, potassium chloride, (beta-D)-Isomer, protein complex, 6-trans-farnesyl-diphosphate diphosphate-lyase [(+)-germacrene-A-forming] activity, 5R)-hexane-1, igaad, medical condition, strontium salt, D-mannitol, mereological quality, [HCl], count in organism, divided_from, Keoflo, water, I2PP2A, Dyrk1, CES2A1, l(3)84Ba, connected anatomical system, ACETONITRILE, tandem MS, Radiations, Temperature, nup32D, 10^[-9], Normality, CC1, Tripcellim, CGI-97, sample population, chromic formate, Carbon 12, Interleukin-1 beta convertase, Protein Gene Products, dSET/TAF-Ibeta, 2610030F17Rik, Pressures, poly(oxyethylene), SSKS, 3.4.22.36, calcium salt, cold (chronic obstructive lung disease), Chronic Obstructive Lung Disease, Runnings, Scl, presence or absence in organism, sodium chloride, Methane, perfluoroethylene, Size, PLIP, IL1BC, MeCN, cadmium salt, Scx, muriate of potash, fond, TFE, D-(-)-Mannitol, bHLHa48, BOUND WATER, growth and development, chronic obstructive pulmonary disease (COPD), oxidane, Type 2A-interacting protein, trichloro-, Long Term, 5730420M11Rik, 2-Propanone, Polypeptides, protein polypeptide chains, hydrogen chloride, scx, cobalt(II) formate dihydrate, Methanal, CH3-C#N, hnu, Dextrose, Chronic irreversible airway obstruction, GRP8, GRP7, Macrogol, Il1bc, HTATIP, 3-Propanetriol, average, Genomes, ITF-2b, CAFL - Chronic airflow limitation, proteins, IL-1BC, AI047805, PA1, set, Wasserstoffchlorid, column, cPLA2, disease (COPD), SEF-2, anatomical tube, bHLHa41, sample, Stainless steel, TIP, Klor-con, Radiation, (2R, subdivided, Ethene, GAST, Cyc E, ANAC091, Vitreous, Polyethylene, Putative MAPK-activating protein PM10, H2O, BG:DS07108.3, Dm1, Chlorwasserstoff, Sizes, TIP60, leukemic reticuloendotheliosis, Airflow Obstruction, END, 6-trans-farnesyl-diphosphate diphosphate-lyase (germacrene-A-forming) activity, forked, Amylomaize, GLYCINE-RICH PROTEIN 8, Chronic Obstructive Airways Disease, HVEML, HLA-DR-associated protein II, glycerine, cromium (+3), alpha-hydro-omega-hydroxypoly(oxy-1, alpha, septate, beta Trypsin, Macrogols, TAF-IBETA, F2G1.4, spirit of wood, Drice, PEG, TAF-Ibeta, methyl cyanide, chlorane, DRICE, lead (+2) salt, Glucose Monohydrate, TIP41, PEO, Methyl Alcohol, F10B6_15, Particle Sizes, GLYCEROL, polysulfoethyl A, nickel formate dihydrate, cycline, InChIKey=WSFSSNUMVMOOMR-UHFFFAOYAT, lead formate, Effects, Glucose, chlorure d'hydrogene, fractured, DrIce, DmcyclinE, aluminum salt, CAO - Chronic airflow obstruction, high weight, Kingdom of the Netherlands, splitted from, protein-containing complex, 2-ethanediyl), and RNA binding 1, F10B6.15, ESA1, Ly113, cdi7, Polyethylene Oxides, cyclinE, SEF2-1, heavy, CDI7, CG42273, disease or disorder, 3.4, Normalities, 2310061K05Rik, beta-Trypsin, Polyethylene glycol, Long-Term, (L)-Mannitol, methanoic acid, DmcycE, Streptoverticillium sp., Solution, CD156, Injectable, DmelCG4579, ms(2)zk, 2PP2A, Steel, NEC, Kochsalz, photon, dSET, tetrafluoroethene, TR2, Polyaethylenglykole, Chromatographies, ASP-I2, Anhydrous, constant, caspase 3, BcDNA:LD21772, COAD, Proteins, Expanded PTFE, split, chloridohydrogen, potassium formate, CE-2, Mischung, acetonitrile, chronic, polypeptide, Natriumchlorid, I-2PP2A, Dm I-2, hemorrhaged, MeOH, (COPD), CAL - Chronic airflow limitation, Magnesium chloride anhydrous, lithium formate, Politef, l(2)k02602, postnatal growth, IL-1 beta-converting enzyme, pore-forming toxin activity, CD156a, primary structure of sequence macromolecule, Longterm Effects, ME-IV, Gene Proteins, D-CycE, Polyglycol, DmelCG1028, DRO15DC96Z, nucleobases, [OH2], hydrochloric acid, EXP, MITF-2B, chronic obstructive pulmonary disease, MITF-2A, [H]C([H])=O, Nup32D, IPP2A2, ANT-C, determination, and rna binding 2, postnatal development, Monohydrate, Hydrogenchlorid, (DL)-Isomer, GORE-TEX, extracted material, Teflon, halite, POE, exact), Ccne, diseases, dJ69E11.3, chronic obstructive airways disease NOS (disorder), AntP1, Chainia sp., Min, diseases and disorders, Tetrafluoraethylen, Effect, foton, DmelCG42273, Peptone, long, bases, Formaldehyd, min, SEF2, copper, SCAN, xscleraxis, cloruro sodico, alpha-hydro-omega-hydroxypoly(oxyethylene), Fluon, dulcite, Starch, SOLO DANCERS, Carbon, poly(ethyleneoxide), dihydridooxygen, ANT-P, Sodium Methoxide, glycine-rich RNA-binding protein 8, CHRONIC OBSTRUCTIVE PULMONARY DISEASE, gamma, 3R, [KCl], phapii, rock salt, ice, aqua, extra or missing physical or functional parts, classic hairy cell leukemia, br37, 4R, Programs, pulmonary disease (COPD), TUBE, Ionen, drICE, 2-ethanediol homopolymer, low temperature, chronic obstructive pulmonary disease and allied conditions, HCL, Chronic airway disease, NOS, Carbinol, COPD NOS, carbinol, Mnb, Visible Radiations, parent ion, Chronic airway obstruction, Visible Radiation, growth pattern, cold, 6C, non-developmental growth, HCl, hydrogen hydroxide, E2-2, MASS, gas, Polyglycols, natrii chloridum, magnesium dichloride, organ system, Chronic Airflow Obstruction, METHANOL, Health, l35Dd, precursor ion, polyethylene glycols, D Glucose, Acetamide, surface active agent, carbonium, chronic obstructive airways disease, Polypeptide, obstructive pulmonary disease (COPD), COLD, chronic obstructive lung disease, base, gluco-hexose, other disease, DISEASE (COPD), surfactants, FBN, DMANTPE1, Gene, Methyl, circadian rhythm, presence, l(2)br37, Polyethyleneoxide, Buffer, formic acid, Glycerine, potassium salt, Pulmonary Disease, Arabidopsis NAC domain containing protein 91, Trihydroxypropane, E421, Polytef, zk, E2.2, cloruro de hidrogeno, propane-1, iones, Tcf-4, uniform, chlorure de sodium, Mannitol, methanol, Visible, eau, Chronic Obstructive, non-neoplastic, OCTD, Kohlenstoff, taf-ibeta, FEP, liquid, cromium (+3) salt, AntP, sodium formate, COPD, data, nickel formate, 3H-labeled, 弁 (Japanese, Glyzerin, not elsewhere classified, 3-triol, strontium formate, Polyethylene Glycol, PULM DIS CHRONIC OBSTRUCTIVE, Peptide, Chronic obstructive pulmonary disease finding (finding), Methanol, polyethylene glycol, development, capillary vessel, natural protein, Protein, AI551343, Hydrochloride, sequence, table salt, FORMALIN, GLYCINE-RICH RNA-BINDING PROTEIN 7, Vacuums, CyeE, FORMALDEHYDE, WMS2, cyanomethane, Glycol, chronic obstructive airway disease, 3-Trihydroxypropane, Dops, Photoradiation, l(2)35Dd, Amine, Methylene oxide, LTg, Anhydrous Dextrose, Holland, Trypure, Ion, Polyoxyethylenes, wood spirit, cardinality, TCV-interacting protein, Peptid, assay, ORW1, AA407739, Glycols, growth, CG3938, Glcparent ion, synthesis, Siderochromes., single-organism biosynthetic process, Siderophore, biosynthesis, precursor, precursor ion, bioformation, formation, anabolism, multicellular organismal biosynthetic processIPP2A2, 2-amino-4-(methylsulfanyl)butanoic acid, Cysteine Hydrochloride, False, L Isomer, Gene, protein, protein-containing complex, PHAPII, 5730420M11Rik, peptide, Polypeptides, L-Isomer Methionine, method, protein polypeptide chains, Methionine, peptido, ClvPrd, template-activating factor I, polypeptide chain, method used in an experiment, Gene Products, L-Methionine, Low, Analysis, Half-Cystine, protein aggregate, Pedameth, SET, proportion, me75, F, peptides, Analyses, Genomes, TAF-I, M, L Cysteine, phosphatase 2A inhibitor I2PP2A, ipp2a2, stubby, 2pp2a, labeling, proteins, D17Mit170, CG10574, T1, DmelCG4299, set, IGAAD, 2PP2A, 2-amino-4-(methylthio)butanoic acid, DmelCG10574, taf-ibeta, dSET, dSet, peptidos, s, Half Cystine, metionina, Racemethionine, ratio, phapii, cou, DL-Methionine, Zinc Cysteinate, protein complex, Proteins, igaad, 2-Amino-4-(methylthio)butyric acid, StF-IT-1, L-Isomer, Tl3, Tl2, Peptide, polypeptide, shortened, Lr, native protein, natural protein, I-2PP2A, Dm I-2, Protein, I2PP2A, Methionin, median, methionine, Hmet, Met, Data Base, HLA-DR-associated protein II, DI-2, I-2Dm, proportionality, rate, L-Cysteine, CG4299, inhibitor of granzyme A-activated DNase, I-2PP1, plan specification, Protein Gene Products, Gene Proteins, dSET/TAF-Ibeta, 2610030F17Rik, TAF-IBETA, Data, alpha-amino-gamma-methylmercaptobutyric acid, quotient, Bra, Peptid, TAF-Ibeta, Polypeptide, AA407739, short, i2pp2a, Liquimeth, Peptide., Data Analysesexplosives chemical, Actinomyces, grupo, bioformation, multicellular organismal biosynthetic process., Actinobacteria Stackebrandt et al. 1997, determination, Orphaned Child, Peptidomics, explosives, Keratoactinomycosis, number, Explosive, actinomycotic madura foot, biosynthesis, infections, Madura foot due to Actinomadura, presence, boil, group, Actinomycotic madura foot (disorder), Orphans, count in organism, explosive compound, chemical analysis, explosives chemicals, Explosives, explosive material, actinomycotic mycetema, tandem MS, anaerobic Actinomyces infection, Actinomycotic mycetema, single-organism biosynthetic process, Genomes, formation, MS/MS, MS2, anabolism, Orphaned, actinomycotic infection, Actinomyces infection, Actinomycotic mycetoma of foot, E430039A18Rik, Rest, Siderophore, INSDC_feature:gene, actinomycetoma, CD156a, Children, CD156, canaliculitis, Actinomycetia, synthesis, Orphan, metastatic, actinomycosis, Actinomycosis, high G+C Gram-positive bacteria, Orphaned Children, Agents, actinomycotic mycetoma of foot, grupos, Actinomyces israeli, assay, quantitative, explosive compounds, Actinomycetes, Madura foot due to Actinomadura (disorder), groupe, Siderochromes, Gruppe, presence or absence in organismOrnithine Sulfate (1:1), explosives chemical, bioformation, Actinobacteria Stackebrandt et al. 1997, conformation, Orphaned Child, Fanconi anemia complementation group E, protein, biosynthesis, Spectrum Analyses, precursor, (DL)-Isomer, protein-containing complex, boil, metabolite levels, Actinomycotic madura foot (disorder), peptide, Polypeptides, Orphans, protein polypeptide chains, ClvPrd, peptido, polypeptide chain, explosive compound, Fanconi anemia complementation group type E, Mass, 5-Diaminopentanoic acid, explosives chemicals, 2, Analysis, protein aggregate, Explosives, explosive material, actinomycotic mycetema, primary metabolism, Mass Spectroscopy, multicellular organismal biosynthetic process, Mass Spectrum Analysis, anaerobic Actinomyces infection, Fanconi anaemia complementation group type E, Synthetic, single-organism biosynthetic process, Fanconi Anemia, peptides, Genomes, Analyses, Biology, formation, anabolism, Orphaned, Actinomyces infection, complementation group E, ornithine, Siderophore, proteins, Ornithine Monohydrobromide, canaliculitis, Actinomycetia, synthesis, Actinomycosis, Orphaned Children, actinomycotic mycetoma of foot, grupos, FANCE, face, peptidos, explosive compounds, relational structural quality, Fanconi anemia caused by mutation in FANCE, Madura foot due to Actinomadura (disorder), Siderochromes, FACE, close to, Fanconi anemia, visage, Actinomyces, grupo, explosives, Keratoactinomycosis, protein complex, facia/facies, Biologies, Explosive, actinomycotic madura foot, Ornithine Monoacetate, infections, Fanconi anaemia caused by mutation in FANCE, Madura foot due to Actinomadura, Spectrum Analysis, Peptide, Ornithine Dihydrochloride, Faces, group, Spectroscopy, near to, polypeptide, 5-Diaminopentanoic Acid, metabolite traits, DL-Ornithine, native protein, natural protein, Operons, Ornithine Hydrochloride, Protein, LCE, 5-Diaminovaleric acid, Ornithine Phosphate (1:1), Mass Spectrum Analyses, Ornithine, parent ion, Mass Spectrum, Actinomycotic mycetema, complementation group type E, Orn, actinomycotic infection, Actinomycotic mycetoma of foot, Rest, Spectrometry, actinomycetoma, INSDC_feature:gene, Ornithine Monohydrochloride, Children, (D)-Isomer, Orphan, actinomycosis, high G+C Gram-positive bacteria, Agents, precursor ion, FAE, approaches, Actinomyces israeli, (L)-Isomer, 5-diaminopentanoic acid, Synthetic Biologies, vicinity of., Peptid, Polypeptide, FANCE Fanconi anaemia, Actinomycetes, groupe, Gruppe, 5 Diaminopentanoic Acid, FANCE Fanconi anemiaSiderochromes., Siderophore, assay, determination, Peptidomics, chemical analysis1701trueProteomics analysis of streptomyces sp. MBT76 demonstrates the production of qinichelins, new mixed catecholate-hydroxamate siderophoresActinomycete genomes contain a plethora of orphan gene clusters encoding unknown secondary metabolites, and representing a huge unexploited pool of chemical diversity. The explosive increase in genome sequencing and the massive advance of bioinformatic tools have revolutionized the rationale for natural product discovery from actinomycetes. In this context, we applied a genome mining approach to discover a group of unique catecholate-hydroxamate siderophores termed as qinichelins from Streptomyces sp. MBT76. Quantitative proteomics statistically correlated a gene cluster of interest (qch) to its unknown chemotype (qinichelin), after which structural elucidation of isolated qinichelin was assisted by bioinformatics analysis and verified by MS2 and NMR experiments. Strikingly, intertwined functional crosstalk among four separately located gene clusters was implicated in the biosynthesis of qinichelins.2018-10-242017-05-24PXD0065774466449917558258064609938804333029535851114512067811080612461962461972842188090320637522995811363350906836351524260275838538791187947398007186801NCBITaxon:1280572307688881114301187263555571468351452627412583649117571508472746707364113546138855821803582329031388869152167786889539978447512951398704351123294126246942229692812397713311809933995232310343043654813583334607925198183659219813143205971293818826826558194762149363295726373719222212502321979116938197912NCBITaxon:1272111957741247818661328388281395584345135867221755437294914081694595418045416193426683413903634530586112041623264460711210411706127443210859561496367469412831666943694559729452811320810039241065837514633087936944784475174150808206542208964317447127020896344597413797912814128384119466940066714545879329935293201019812646904106611457551216599404833925121659721659545501315282697316062121331283128212803395232181083232178252529204353152277944762376281123531534543498257419481478717203094980197525556978153623142410439841356742951053948824261914548140479NCBITaxon:2157115062155153129243989903712285778299659753205968762319351630980058884558697818178081736309431791036905660698936160488566140985910478256641624251505907118702348211870532049138169363063863235554320466322261866331299456586068845351512623217219217459216129567171360441001602358700264203694569410167634102107612773511316582150480239254727933216042814610029135843035786174633132529111870021576418096997566545774449156666754760445246767568403019172651813019411192943963366411363367217511176451023987093202533114NCBITaxon:40559325823590927883965236526498217870749821634725656915693872249821165448720569734749516529981666905108944835525663661410NCBITaxon:4932871556891758543102459808108944475741755518100058945268087322875351908323311561263730069303911415542193255521872587268723872487298727123598755529NCBITaxon:475154762422314146547730551800664417725564843376771581419198822445299767446982510216998231174673897641041051155744689444474153NCBITaxon:5055796016776744685100036487529600NCBITaxon:58116534911822637082483341475480557841946751299114578458359741276473194439137086326547876569835566369838192011164034624136810893161001428099612NCBITaxon:548681NCBITaxon:582054641498523077479464098609958652283637379196627290587668960596061572951372218759446708756227086266596496041711013354833090107687834974129829986038783481062675664306151279365431947470334249472235985961089615886559746499870392021110728782721540782119085037570752983511708282061892577714804189854873502471587796273002515630411902528674636046147964696443759394911462472746128746011345062532566484526591972219638191552638177420858859652639NCBITaxon:615750521028307989511051723750595761313024097200026341961052641109094694077227746941634856899611238694975157244NCBITaxon:2NCBITaxon:1352317036125061747679274506272491596153840052573132464321368615059626979686662990346941713443463603475153632964404210181943010867310185109760635586331496161286287246200966927294227294351338132169685835518653685959689128017260710185687968129411009118696141633316780781186941436183NCBITaxon:6191118698840231719475932118691443906NCBITaxon:102982100071257118348824387279694836419080218787864515970384766425731055347095145122578656870861822607046183260705260707NCBITaxon:7803906NCBITaxon:1502451516658457702921158612184519307920928520131974707856100713370565295076192126385454195NCBITaxon:3055504457391538337991399878168192875NCBITaxon:10239NCBITaxon:7142829126806315327341478281309570547044830064175158592381055524159084128108228233446900815025754122810451930174830085210977970671356225833136185509924NCBITaxon:56938268830640527796179469958334240906151015151034809NCBITaxon:63124067448242813076707087613837047592723708281677285035158199011951953370249240536088817562615835583337602126719285321773572671193501846451772998086167879177437371110490563712153481724073714121532395767690147266470913170475712562745790311333550372885962859634792491308100787126387121203511082332648373153214693NCBITaxon:961538323594633185862607995837014126220029813141313493760833344926701311833331114970192812396553972833325283010184138569591110065812375612750454266625515197887492003750360094129349715300910766204114902801483524725175014012573983649908515151438992NCBITaxon:1155237453988374744544641521399274356235931179286643235983906288705158059553611945993271602938511497861603293338922853422999810894535936457428412754121536024271831627713512251179948875058NCBITaxon:1040724271550277916797181673871349130627415895521010306502780291595412629158660025158451584937653271595702100226376733889423451287689135588675512930945146479203124216413310515280382803519284343690935815398580894621080348179392118259028788929502711314560990690174934681927256335442273218903829286757227901107601247190400727NCBITaxon:579653745711431123352356211402064119810871355477142450722953379698355522221291634708714104577512621611542931575461396224213929928713907702020603281857323114319338293550833292307411292211481640278489616010677252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