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Samples were subsequently reduced and alkylated with 5 mM TCEP and 20 mM CAA at 37 °C for 1 h. LysC was added in an enzyme:protein ratio of 1:200 (wt/wt) and incubated with the sample for 2 h at 37 °C. After dilution of the sample to 2 M Urea, Trypsin was added at an enzyme:protein ration of 1:100 (w/w) to digest overnight at 37 °C. Trypsin was quenched with 1% FA and centrifuged for 10 min at 20,000 x g. The supernatant was collected and desalted using C18 StageTips and lyophilized. Peptides were quantified using BCA peptide quantification (Thermo Fisher Scientific). Peptide mixtures after tryptic digest were analyzed by a reversed-phase capillary liquid chromatography system (Dionex Ultimate 3000 NCS-3500RS Nano, Thermo Fisher Scientific) connected to an Orbitrap Fusion mass spectrometer (Thermo Fisher Scientific). For sample loading, a PepMap C18 trap-column (Thermo Fisher Scientific) of 75 µm ID x 50 mm length, 3 μm particle size, 100 Å pore size was used. The loading mobile phase A contained 1% acetonitrile and 0.1% TFA acid in water, and mobile phase B 0.1% TFA acid in acetonitrile. Reversed-phase separation was performed using a 50 cm analytical column (in-house packed with Poroshell 120 EC-C18, 2.7 µm, Agilent Technologies) with mobile phase A containing 0.1% formic acid in water, and mobile phase B 0.1% formic acid in acetonitrile. The gradient started with 4% buffer B reaching 80% buffer B, with total run time of 80 or 120 min including column wash and equilibration at a flow rate of 250 nL/min. For standard protein identification and quantification, MS1 scans were acquired in a range of 375 to 1500 m/z with 50 ms max. injection time, a resolution of 120,000 and an AGC target value of 4e5. Precursor ions with charge states 2-4 were isolated with a mass selecting quadrupole (isolation window m/z 1.6) with 40 s dynamic exclusion. MS2 scans were recorded with a linear ion trap in rapid mode with dynamic maximum injection time and mass range, standard AGC target and an NCE of 30% (1 s duty cycles). For pyrophosphorylation studies, settings were kept the same apart from the following changes: MS1 scans were recorded with a mass range of 380 to 1400 m/z and isolated with a dynamic exclusion of 15 s. Precursors were fragmented with 25% CID and recorded in the Orbitrap at a resolution of 15,000 with 100 ms maximum injection time and 1e4 AGC target. If a targeted loss of 177.9432 Da (± 25 ppm) was detected, the precursor was again isolated and fragmented in the MS2 with 30% EThcD, 2000 ms max. injection time, 1e5 AGC target, at a resolution of 120,000 in a m/z range of 200–3000 (2-3 s duty cycles). For phosphorylation studies, no prior CID-MS2 scans and targeted loss trigger were performed and all precursors have directly been fragmented by EThcD."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Raw data were analyzed and processed using MaxQuant. Protein identification was performed with default settings. Search parameters included two missed tryptic cleavage sites, fixed cysteine carbamidomethyl modification, and variable methionine oxidation and N-terminal protein acetylation. Label-free quantification and match between runs were enabled. Database search was performed against the Human UniProt/Swiss-Prot database with common contaminants. The false discovery rate (FDR) was set to 1% at both peptide and protein level. The analysis of the quantitative proteomic data was carried out using Perseus.   7 Proteins were filtered to exclude reverse database hits, potential contaminants, and proteins only identified by site. Proteins were further filtered by rows, requiring valid values for at least two out of four technical replicates. Data was imputed using Perseus default parameters. Volcano plots were generated using a t test (number of randomizations: 250)."],"omics_type":["Proteomics"],"labhead":["Dorothea Fiedler"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Leibniz-Forschungsinstitut fuer Molekulare Pharmakologie Humboldt-Universitaet zu Berlin"],"species":["Homo Sapiens (human)"],"publication":["10.1021/JACS.4C03069"],"submitter_mail":["ruwolt@fmp-berlin.de"],"submitter_affiliation":["Leibniz-Forschungsinstitut for Molecular Pharmacology"],"submitter_country":["Germany"],"sample_synonyms":["Water, sodium salt, l(4)17, l(4)13, ammonium formate, Gli, Size, 13C-labeled, MeCN, cadmium salt, dmMOF, ion, TFA, Cid[Mel], Ci[D], Particle, H(2)O, BOUND WATER, Basodexan, Capillary, magnesium formate, nutcracker syndrome, X-50, oxidane, CG3629, zinc salt, CENP-A, phosphorylation, RP11-508D10.1, Long Term, WATER, CENP-C, HOH, Bca, l(1)AA33, DmelCG9648, Polypeptides, cobalt(II) formate dihydrate, Beds, CH3-C#N, dMOF, B1, 1, 2, AGM4, 3, 4, 9, WMS, bca, type 5 acid phosphatase, pore, BcDNA:RE21270, DmelCG13176, weight-weight percentage, ORF19, T6G21.3, Sinusoidal Bed, Ba, AI047805, Bed, T-cell antigen Gp39, 15-disilaoctadecane, CI, column, l(1)B2/13.1, sample, D1, EThcD, TNFSF5, nickel salt, SGS, CiD, Ce, l(1)LB9, Stars, Ci, dl, cobalt (+2) salt, Tnfrsf5, acid, familial, Liquid Chromatography, Longterm Effect, Sinusoids, not genetically inherited, collisionally activated dissociation, Cid, CID, ciD, hCD40L, bHLHd7, ACMICD, bHLHd6, l(2)01092, bHLHd9, bHLHd8, H2O, 11-tetrathia-4, sodium (4:1:1) salt, bHLHd5, bHLHd4, Dm1, magnesium salt, IMD3, Sizes, ci-D, 1110049F14Rik, Hydrogen Oxide, Acid, DLL, CENP-A/Cid, CENP-A/CID, LY57, CENPA, NKTL, formate, cromium (+3), beta Trypsin, E(Arp), w/w, dll, Charges and Fees, ammonium (4:1) salt, methyl cyanide, lead (+2) salt, STARS, Ci-155, Particle Sizes, nickel formate dihydrate, lead formate, Effects, Biocatalysts, Ly62, fractured, TAPK, aluminum salt, Carbamide, precursor, protein-containing complex, thomson, TR-AP, CG7826, agua, mass-to-charge ratio, CenpA, DmelCG3025, l(2)387, Ly57, GP39, Gene Products, CG7835, CG42273, Capillary Bed, Carmol, Siah, NCS, gp39, CD154, MAX, anatomical systems, FAM39E, CATC4, GKLP, thallium (+1) salt, Longterm, Sinusoidal Beds, beta-Trypsin, rubidium salt, Long-Term, methanoic acid, ions, EThcD., CD156, Injectable, PCTAIRE2-binding protein, Enzyme, max, MGC:45012, CG2125, peptidos, Saeure, 2.7, PTHB1, LB5, NTKL, mass percentage, 1-(14)C-labeled, Phase Separations, Proteins, CG3025, Phosphorylations, split, RNS, potassium formate, copper (+2) salt, backward, ACP5, Rnt, acetonitrile, SPENCDI, buffer, CD142, lLB5, polypeptide, Wash2, Wash1, native protein, carbonyldiamide, ci[D], C18, cupric formate, Long Term Effects, hemorrhaged, MFS1, Sinusoidal, aluminum formate, AU020952, torn, Phase, lithium formate, DmelCG2125, CenH3, ci155, underdeveloped, Saeuren, Separations, Striated muscle activator of Rho-dependent signaling, pore-forming toxin activity, ethanenitrile, CD156a, Longterm Effects, ME-IV, Gene Proteins, Capillary Beds, Left renal vein entrapment syndrome, Fees, TRACP, [OH2], CenpA/CID, reversed, channel-forming toxin activity, NCMe, leg, zinc formate, lead salt, protein, 10, peptide, Tnfsf5, 15, ammonium tetraformate, peptido, Min, protein aggregate, Effect, Injectables, DmelCG42273, T-BAM, CG13329, Art, tartrate-resistant acid ATPase, peptides, E927b, dMax, E430039A18Rik, hypoplasia, min, mAPC, copper, genetic, TRAP3, Tudor repeat associator with PCTAIRE-2, lithium salt, RUN, Run, 380, ppm, Cenp-A, dihydridooxygen, TF, TEAB, Lyw-57, blood capillary, T5ap, dihydrogen oxide, MGC - 45012, GPHYSD2, ammonium (2:1) salt, Long-Term Effects, Sinusoid, CD40L, Cd40l, aqua, Dmel_CG7826, CG9648, Th, fragmented, ur, AA33, SLP65, carbamide, Ly-57, Renal Nutcracker Phenomenon, Ionen, DmelCG3629, dmax, BcDNA:LP01770, TNF-related activation protein, WASH, nickel (+2) salt, Ly-62, Dmel_CG7835, 15-tetraethoxy-, cenpA, Mnb, MNB, parent ion, SLP-65, ammonium salt, CenH3/CID, Ci/Gli, Ci/GLI, BASH, MS/MS, acide, cracked, hydrogen hydroxide, cobaltous formate, acids, AW743063, acido, MASS, AW124434, AI326936, En(Arp), acqua, loss of, H2NC(O)NH2, MOF, Mof, precursor ion, Charge, TrATPase, IGM, copper salt, Biocatalyst, CD40LG, Polypeptide, Wasser, 1728, CG1849, TRAP, p50, cesium salt, FBN, Gene, 2-(1-benzyl-4-methylpyrazol-3-yloxy)-2-methylpropionic acid, formic acid, Buffer, Karbamid, l(4)102ABc, potassium salt, reduced, Ms1, mixt. with silica, Injection, ECTOL1, 14C-labeled, p63, Harnstoff, p65, tiny, CD40-L, CenH3[Cid], CenH3[CID], MS1, iones, MS2, Runt, DmelCG13329, eau, calcium formate, Charges, OCTD, X-linked combined immunodeficiency, Fee, 10^[-6], TEIF, Trap, male sterility 1, HIGM1, cromium (+3) salt, Long-Term Effect, sodium formate, TCEP, C78062, BLNK-S, small, P105, DYRK1, nickel formate, 3H-labeled, protein complex, PCTAIRE2BP, TSH1, DmelCG1849, strontium formate, cenH3, Bp50, NY-CO-33, strontium salt, Peptide, AI875693, CAA, capillary vessel, CAD, Ci155, 16-Dioxa-8, water, Protein, Dyrk1, SDCCAG33, ACETONITRILE, tandem MS, cyanomethane, WMS2, CG13176, Separation, PRSS, uree, MALE STERILITY 1 PROTEIN, CC1, Tripcellim, cerebral amyloid angiopathy, l(1)19Ea, sample population, chromic formate, Protein Gene Products, P235, Trypure, Ion, AA960152, SSKS, calcium salt, Peptid, CID/CENP-A"],"name_synonyms":["N-acetyl-D-glucosamine kinase, 2.7.1.59, Activity, protein complex, Proteins, N-acetylglucosamine kinase, Phosphorylations, ATP acetamide-2-deoxy-D-glucose - 6-phosphotransferase, Gene, GlcNAc kinase, protein, protein-containing complex, acetylaminodeoxyglucose kinase, rat., phosphorylation, human, Protein Gene Products, Gene Proteins, native protein, NAGK protein, Protein, Gene Products, protein aggregate, General activity, N-acetylglucosamine kinase protein"],"data_synonyms":["Bru, IPP2A2, Cysteine Hydrochloride, E 920, 2-amino-4-(methylsulfanyl)butanoic acid, human being, Raw, determination, number, cleavage, L-Zystein, L Isomer, Gene, LFQ, protein, protein-containing complex, presence, PHAPII, Human, 5730420M11Rik, peptide, Polypeptides, L-Isomer Methionine, Methionine, peptido, Homo sapiens, Gene Products, L-Methionine, Del(8)44H, 2-amino-3-mercaptopropanoic acid, Cystein, Half-Cystine, protein aggregate, Man, Pedameth, Svc, C, SET, Man (Taxonomy), peptides, TAF-I, cisteina, L Cysteine, M, E-920, protein amino acid acetylation, ipp2a2, 2pp2a, Hcys, CG10574, DmelCG4299, IGAAD, set, 2-amino-4-(methylthio)butanoic acid, 2PP2A, CYSTEINE, DmelCG10574, taf-ibeta, dSET, dSet, FREE CYSTEINE, site, peptidos, Half Cystine, (2R)-2-amino-3-mercaptopropanoic acid, metionina, Racemethionine, phapii, Zinc Cysteinate, DL-Methionine, protein complex, Modern, Proteins, 2-Amino-4-(methylthio)butyric acid, igaad, StF-IT-1, L-Isomer, Cys, Peptide, group, polypeptide, count in organism, native protein, I-2PP2A, Protein, Dm I-2, chemical analysis, I2PP2A, Methionin, Hmet, (2R)-2-amino-3-sulfanylpropanoic acid, fixed, Zystein, region, Met, Data Base, Col4a-1, HLA-DR-associated protein II, ensemble, DI-2, 2-amino-3-sulfanylpropanoic acid, I-2Dm, L-2-Amino-3-mercaptopropionic acid, common, L-Cysteine, (R)-2-amino-3-mercaptopropanoic acid, CG4299, cardinality., 2-Amino-3-mercaptopropionic acid, human, L-Cystein, Protein Gene Products, I-2PP1, Gene Proteins, dSET/TAF-Ibeta, 2610030F17Rik, TAF-IBETA, Modern Man, alpha-amino-gamma-methylmercaptobutyric acid, E920, Peptid, TAF-Ibeta, Polypeptide, assay, variable, AA407739, Liquimeth, i2pp2a"],"description_synonyms":["author summary, Post Translational Amino Acid Modification, HSA242910, Striadyne, posttranslational modification, PhrB photolyase activity, Activity, Adenosine Triphosphate, Dambose, ATP acetamide-2-deoxy-D-glucose - 6-phosphotransferase, protein, Progress Reports, pigmented epithelium, Protein Processing, phosphorylation, Social Controls, 2-Amino-3-hydroxypropionic acid, dmTAF[[II]]230, Meat sugar, rat, Readability, Mesoinositol, i-inositol, Summary Report, 1, 2, 3, 4, 5, NUP96, Kinase, protein aggregate, epithelium, Summary Reports, Formal Social Controls, imprinted and ancient gene protein, Post-Translational Protein, D-myo-Inositol, inosite, posttranslational amino acid modification, GNK, Gnk, Chromium Adenosine Triphosphate, TFIID TAF250, pre-mortem, Progress Report, Post-Translational, cel, L-myo-Inositol, pigmented retina, Posttranslational Protein Processing, DNA cyclobutane dipyrimidine photolyase activity, SUPPRESSOR OF AUXIN RESISTANCE 3, 2R, Social, Progress, set, ATPsyn b, Field Reports, 2-amino-3-hydroxypropanoic acid, CrATP, ATP:2-acetylamino-2-deoxy-D-glucose 6-phosphotransferase activity, Cr(H2O)4 ATP, NAGK protein, ATP Phosphotransferases, ATP, single organism signaling, Chromium Ammonium Salt, 3S, N-acetyl-D-glucosamine kinase, ATPsyn-&bgr, PRE, Post-Translational Modification, dTAF[[II]]230, anatomical protrusion, ATPsyn-b, (1r, 6S)-cyclohexane-1, deoxyribonucleic cyclobutane dipyrimidine photolyase activity, Modifications, N-acetylglucosamine kinase, TAF200, cis-1, TAFII-250, TAF250/230, 6-hexol, TAFII250, Post Translational Modifications, retinal pigment, Investigative Report, 3-Hydroxyalanine, Manganese Salt, dipyrimidine photolyase (photosensitive), Magnesium Salt, retinal pigment layer, region, ATP:N-acetyl-D-glucosamine 6-phosphotransferase activity, 5R, 4s, CG11154, ATP - N-acetyl-D-glucosamine 6-phosphotransferase activity, 1L-myo-Inositol, Transphosphorylases, PTM, 1D-myo-Inositol, Processing, Field, Control, Controls, 5/4, CG17603, TAF[[II]], human, summary, CaATP, living, phr A photolyase activity, meso-Inositol, Report, DNA-photoreactivating enzyme, Amino Acid Modification, Taf250, spine, post-translational modification, SR3-5, Cells, 6-HEXAHYDROXY-CYCLOHEXANE, MgATP, L Serine, Posttranslational Modifications, E430016J11Rik, L-Serine, Regulation, Posttranslational Modification, TAF230, Eukaryotic, Gruppe, Regulations, d230, Post-Translational Protein Modifications, 6-cyclohexanehexol, Magnesium Adenosine Triphosphate, ATP-MgCl2, Gene, dTAFII250, ATP - 2-acetylamino-2-deoxy-D-glucose 6-phosphotransferase activity, photoreactivating enzyme activity, IB, protein-containing complex, EfW1, Ximpact, Adenylpyrophosphate, Posttranslational, protrusion, protein amino acid phosphorylation, Investigative, dmTAF1, Taf230, ATP-synbeta, Gene Products, Post Translational Modification, Post-Translational Modifications, stratum pigmentosa retinae, acetylaminodeoxyglucokinase activity, Phosphotransferase, Post Translational, Cyclohexitol, TAF250, Modification, study, Chiro Inositol, Taf200, MOS3, deoxyribocyclobutadipyrimidine pyrimidine-lyase activity, dTAF[[II]]250, PRECOCIOUS, DmelCG11154, cell, beta-ATPase, ATP MgCl2, F23A5.3, Taf1p, posttranslational protein modification, Transphosphorylase, acetylaminodeoxyglucose kinase, Protein Modifications, Post-Translational Amino Acid Modification, dTAF250, acetylglucosamine kinase (phosphorylating), Chromium Salt, ATPasebeta, Posttranslational Amino Acid Modification, grupos, synopsis, MnATP, ATPase beta, Bios I, Posttranslational Protein, site, TAF, Investigative Reports, ATPB, F23A5_3, N-acetylglucosamine kinase protein, TAF[[II]]250, grupo, Formal Social Control, protein complex, Proteins, 5-trans-4, Phosphorylations, Post-Translational Protein Modification, Magnesium Chloride, l(3)84Ab, 2-acetylamino-2-deoxy-D-glucose kinase activity, BG:DS00004.13, function, Adenosine 5'-(tetrahydrogen triphosphate), Cell, impact-a, Phosphotransferases, group, dTAF230, ATP-syn-B, Inositol, Serin, MODIFIER OF SNC1, Atriphos, ML-1, native protein, Social Control, p230, Protein, Research Reports, deoxyribonucleic photolyase activity, TAF[[II]]250/230, TFIID, IMPACT, imprinted and ancient gene protein homolog, Kinases., Protein Modification, Taf[[II]]250, RPE, Manganese Adenosine Triphosphate, TAF[[II]]230, 2.7.1.59, distinct, ensemble, photolyase activity, Post Translational Protein Processing, Ins, Chiro-Inositol, Myoinositol, Rest, TAF[II]250, GlcNAc kinase, Understanding, post-translational amino acid modification, p. pigmentosa retinae, Protein Gene Products, Eukaryotic Cell, Gene Proteins, inositols, Post Translational Protein Modification, DmelCG17603, signalling process, Reports, ATPIB, Post-Translational Protein Processing, GlcNAc kinase activity, deoxyribonucleate pyrimidine dimer lyase (photosensitive), regulation, Summary, groupe, General activity, RWDD5, Field Report, Calcium Salt, TAF1"],"additional_accession":[]},"is_claimable":false,"name":"An Uncommon Phosphorylation Mode Regulates the Activity and Protein Interactions of N-Acetylglucosamine Kinase","description":"Reversible protein phosphorylation serves as a pivotal signaling mechanism in eukaryotic cells, contrasting with the poorly understood protein pyrophosphorylation, a posttranslational modification (PTM) whose functions in living organisms remain elusive. Unlike kinase-mediated protein phosphorylation, pyrophosphorylation is thought to be installed non-enzymatically, when a highly phosphorylated inositol pyrophosphate (PP-InsPs) transfers its high energy β-phosphoryl group onto the pre-existing phosphoryl group of a protein substrate. Our group recently presented the initial evidence of endogenous protein pyrophosphorylation in mammalian cell lines, including N-acetylglucosamine kinase (NAGK). Here we report, the first site-selective pyrophosphorylation of NAGK, strategically modified at its endogenous site to evaluate the modification's impact on protein activity. Biochemical evaluation revealed a significant reduction in GlcNAc kinase activity upon phosphorylation and near-complete inactivation upon pyrophosphorylation at serine 76 (S76), highlighting pyrophosphorylation’s unique regulatory function facilitated by an ATP-mediated autocatalytic process. ppS76-NAGK demonstrated notable stability in mammalian cell lysates, hinting at alternative functional outcomes. Proteomic examination unveiled distinct protein-protein interactions for ppS76-NAGK, suggesting involvement in diverse cellular pathways. In summary, this study enhances our comprehension of protein pyrophosphorylation and its significance for kinase regulation and protein-protein interactions, paving the way for further exploration of pyrophosphorylated kinases.","dates":{"publication":"2024-05-22","submission":"2024-02-12"},"accession":"PXD049339","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:112509","NEWT:475174","NEWT:111205","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:2736291","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:294671","NEWT:376619","NEWT:1748418","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:1907535","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:166340","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:3357","NEWT:111225","NEWT:7719","NEWT:29073","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:2767846","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:468822","NEWT:1912919","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:37919","NEWT:342808","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:709964","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:156974","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:1538","NEWT:2065263","NEWT:199225","NEWT:177437","NEWT:717966","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:29022","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:1883426","NEWT:8165","NEWT:96794","NEWT:10401","NEWT:10886","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:2714896","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:1608994","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2709072","NEWT:2499525","NEWT:1122981","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:141469","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:1922344","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:279010","NEWT:291146","NEWT:29466","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:911022","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:717989","NEWT:33952","NEWT:1255232","NEWT:11723","NEWT:445","NEWT:153009","NEWT:1004879","NEWT:261756","NEWT:166314","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:1871049","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:156981","NEWT:30455","NEWT:28141","NEWT:9031","NEWT:2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