Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110N_1.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_1.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110P_1.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_DIIOV_K151L_1.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_K151L_1.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_WT_1-1.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K151L_1.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K113A_1_220724.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110V_2.msfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/checksum.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110P_1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110V_2.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110N_1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_WT_1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K151L_1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_DIIOV_K151L_1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_K151L_1.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K113A_1_220724.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K113A_1_220724.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_DIIOV_K151L_1.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_K151L_1.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_1.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110P_1.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110V_2.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_WT_1-1.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K151L_1.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110N_1.mgfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_DIIOV_K151L_1.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K113A_1_220724.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_WT_1-1.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_K151L_1.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110P_1.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110V_2.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110N_1.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K151L_1.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_1.mzTabftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_WT_1-1.pdResultftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110N_1.pdResultftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_1.pdResultftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_Y158F_K151L_1.pdResultftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K151L_1.pdResultftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110V_2.pdResultftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_K113A_1_220724.pdResultftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_DIIOV_K151L_1.pdResultftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD075155/MjGATase_D110P_1.pdResultprimary200OKhb@jncasr.ac.inHemalatha BalaramMass SpectrometryBottom-up proteomicsLc-ms/msDeamidationSuccinimidehttps://www.ebi.ac.uk/pride/archive/projects/PXD075155The protocol followed was as described earlier (Kumar et al., 2016, Nat commun) and derived from that developed by Mann and co-workers (Shevchenko et al., 2006, Nat. Protoc). Briefly, 30 μg of the protein was run on 12% SDS-PAGE and stained with Coomassie Brilliant Blue. The stained gel piece carrying the protein band was cut into small pieces, washed with water, and destained using a 1:1 ratio of methanol and 50 mM Tris HCl, pH 7.0. The gel pieces were dehydrated by first using a 1:1 ratio of acetonitrile and 50 mM Tris HCl, pH 7.0, followed by 100% acetonitrile for 30 seconds. The liquid was removed, and the gel pieces were dried in speed-vac and rehydrated in 25 mM dithiothreitol in 50 mM Tris HCl, pH 7.0, and incubated at 56 ℃ for 20 minutes. Thereafter, the gel pieces were washed with water, dehydrated, and dried as described above. The gel pieces were rehydrated with buffer containing 500 ng of trypsin in 50 mM Tris HCl, pH 7.0 such that it covered the gel pieces and incubated at 37 °C for 12 hours. The tryptic peptides were extracted twice with 50% acetonitrile and 5% formic acid. The extracted peptides were pooled together and dried in a SpeedVac. The dried peptides were dissolved in 2% acetonitrile and 0.1% formic acid and desalted using C18 spin columns (Thermo Scientific). The fractions eluted from the C18 spin columns were dried using a SpeedVac. The dried peptides were dissolved in 20 μl of MS-grade water with 0.1% formic acid. Mass spectral analyses were carried out on a Q Exactive HF mass spectrometer (Thermo Scientific, USA) equipped with an Easy nanoLC 1200 (Thermo Scientific). Data were acquired in positive ion mode using the HCD fragmentation method. Easy-Spray column (Thermo Fisher Scientific), PepMap RSLC C18 of 2 μm particle size, 100 Å pore size, 75 μm inner diameter and 25 cm length along with a C18 guard column (Acclaim PepMap100), of 3 µm particle size, 100 Å pore size, 75 µm inner diameter and 2 cm in length was used. Column temperature was set to 40 ℃ and autosampler temperature was set to 4 ℃. Water with 0.1% formic acid as solvent A and 80% acetonitrile with 0.1% formic acid as solvent B was used. The flow rate was set at 300 nl min-1. The peptides of the MjGATase_SNN109 and mutants were eluted using the following steps; a gradient of 5% B to 25% B in 65 mins, isocratic at 25% B for 5 mins followed by a gradient of 25% B to 95% B over 15 mins, isocratic at 95% B for 10 mins, gradient of 95% B to 5% B in 2 mins and finally equilibration at 5% B for 8 mins. The mass spectrometer was operated in a data-dependent mode with a scan range of 300 – 3800 m/z and a resolution of 120,000 at m/z 200 Th. The top 15 abundant ions were fragmented by HCD at a normalized collision energy of 30 and the ddMS2 was acquired at a resolution of 15,000 at m/z 200 Th, a scan range of 200 – 2000 m/z, and dynamic exclusion set to 5 sec. The ion injection time and ion target value were set at a threshold of 50 ms and 3e6 for MS scans and 50 ms and 1e5 for MS/MS scans. A nanospray ion source was used with a spray voltage of 1.7 kV, capillary temperature of 275 ℃, and S-lens RF level set to 55. Data acquisition was carried out using Xcalibur software v.4.1 (Thermo Scientific).PrideNot availableData analyses were done with Proteome Discoverer (PD) v.2.3. and v.2.4. software and Qual browser of Xcalibur software v.4.1 (Thermo Scientific). The search engine, Sequest HT in PD was used with an input of MjGATase sequence into the protein database. The following search parameters were used: precursor mass tolerance set to 10 ppm and fragment mass tolerance of 0.05 Da. The maximum missed cleavage sites allowed for tryptic peptides was set to 2. Dynamic modifications of oxidation at Met (+15.995 Da), ammonia-loss at Asn (-17.027 Da) and deamidation at Asn/Gln (+0.984 Da) leading to the conversion of -CONH2 to -COOH were included.ProteomicsHemalatha BalaramMolecular Biology and Genetics Unit, Jawaharlal Nehru Centrefor Advanced Scientific Research, Bengaluru, IndiaCOMPLETEMethanocaldococcus Jannaschiihb@jncasr.ac.in42055332 Chandrashekarmath A, Jash O, Singh K, Bellur A, Roy CS, Dongre A, Chathoth NE, Anjukandi P, Kumar S, Mukherjee S, Balaram P, Balasubramanian S, Balaram H. Insights into the role of internal catalysts in asparagine-to-succinimide conversion in a hyperthermophilic glutamine amidotransferase. J Biol Chem. 2026 302(6):113093 10.1016/j.jbc.2026.113093ProfessorIndia10.6019/PXD075155Succinimide (SNN), an intermediate spontaneously formed during asparaginyl deamidation or aspartyl dehydration in proteins, is generally hydrolysis-prone, leading to isomerization to an L/D α/β-aspartyl residue, with the latter being considered deleterious to protein structure and function. An unusually stable SNN-mediated conformational rigidity through restriction of the backbone dihedral angle, ψ, enhances the thermostability of glutamine amidotransferase (GATase) from Methanocaldococcus jannaschii (Mj). Although several structural features involved in maintaining a stable SNN and imparting SNN-mediated thermostability have been identified in MjGATase, the residues in the protein that catalyse the rapid and complete conversion of Asn109 to SNN remain unknown. Here, we investigated several mutants of MjGATase for their ability to retain the Asn109 side chain in the unmodified form. Mass spectrometric analysis of 10 single amino acid variants enabled the identification of residues that impacted the proportion of SNN and Asn population in the protein sample. This led to the generation of two double mutants of MjGATase that retained an intact Asn109 side chain, as confirmed by mass spectrometry and crystal structure analysis. These mutant proteins with intact Asn residue at position 109 displayed lower thermal stability than the protein with the SNN modification. Further understanding of the deprotonation mechanism was addressed using quantum mechanics/molecular mechanics molecular dynamics metadynamics simulations.Insights into the role of internal catalysts in asparagine-to-succinimide conversion in a hyperthermophilic glutamine amidotransferase.Chandrashekarmath Anusha A, Jash Oishika O, Singh Karandeep K, Bellur Asutosh A, Roy Chitralekha Sen CS, Dongre Aparna A, Chathoth Nayana Edavan NE, Anjukandi Padmesh P, Kumar Sanjeev S, Mukherjee Souradip S, Balaram Padmanabhan P, Balasubramanian Sundaram S, Balaram Hemalatha HfalseMS/MS analysis of in-gel tryptic peptides of wild-type and mutants of glutamine amidotransferase from Methanocaldococcus jannaschiiA stable post-translational modification, succinimide (SNN), found in Methanocaldococcus jannaschii (Mj) glutamine amidotransferase (GATase) is shown to impart hyper-thermostability to the protein. To explore the role of neighbouring residues in enabling deamidation of Asn109 to SNN, intact protein mass analysis on 12 mutants of MjGATase was performed. Among these mutants, 8 showed varied proportions of mass corresponding to unmodified Asn109. SNN is known to undergo rapid hydrolysis to Asp/iso-Asp, hence, this population can also exist. As Asn intact and Asp/iso-Asp populations differ by only 1 Da, ambiguity exists in their assignment. Therefore, to determine the true levels of unmodified Asn109 population, in-gel trypsin digestion followed by MS/MS analysis of the tryptic peptides for 8 mutants and wild-type of MjGATase was conducted. The MS/MS analysis of two double mutants assured that they indeed retain Asn109 intact, forming a major population. This study points toward the presence of internal catalysts that enable the deamidation of Asn109 to SNN in MjGATase.2026-06-082026-03-03PXD075155NEWT:3555NEWT:71647NEWT:330879NEWT:260201NEWT:35554NCBITaxon:12939NEWT:32046NEWT:544496NEWT:59529NEWT:2042546NEWT:32049NEWT:1392696NEWT:259447NEWT:295546NCBITaxon:2719036NEWT:45351NEWT:445974NEWT:43179NEWT:180454NEWT:5722NCBITaxon:1280NEWT:112503NEWT:1129NEWT:309807NEWT:10245NEWT:55153NEWT:89184NEWT:309800NEWT:281395NEWT:1211601NEWT:876138NEWT:44271NEWT:193516NEWT:351581NEWT:43186NEWT:428146NEWT:111205NEWT:1117NEWT:498257NEWT:10036NEWT:1590NEWT:498019NEWT:1351NEWT:1438992NEWT:1352NEWT:2649997NEWT:1147161NEWT:661410NEWT:638632NEWT:263737NEWT:224326NEWT:1333499NEWT:376619NCBITaxon:79857NEWT:1096976NEWT:5702NEWT:95648NEWT:1589NEWT:135622NEWT:1349NEWT:35786NEWT:1580NEWT:399784NEWT:96731NEWT:383379NEWT:1401257NEWT:418106NEWT:10029NEWT:913645NEWT:44491NEWT:641809NEWT:27933NEWT:556484NEWT:317447NEWT:4688NEWT:7955NEWT:111225NEWT:7719NEWT:7959NEWT:2261NEWT:868565NEWT:94323NEWT:31156NEWT:3112NEWT:79329NEWT:4442NEWT:192875NEWT:1214915NEWT:30069NEWT:12637NEWT:59729NEWT:2164133NEWT:284218NEWT:295105NEWT:108061NEWT:60711NEWT:1316931NEWT:224308NEWT:3347NEWT:511145NEWT:160621NEWT:212790NEWT:931281NEWT:4432NEWT:5762NEWT:5763NEWT:658457NEWT:1310161NEWT:77133NEWT:295358NEWT:145481NEWT:1310165NEWT:29058NCBITaxon:79824NEWT:2246NEWT:2652724NEWT:5757NEWT:68415NCBITaxon:4563NEWT:5755NEWT:44688NEWT:44689NEWT:44447NEWT:752555NEWT:498211NEWT:5759NEWT:347256NEWT:1736231NEWT:5518NEWT:398007NEWT:1527468NEWT:1392NEWT:498217NEWT:498216NEWT:2242NEWT:11320NEWT:286NEWT:391619NEWT:246196NEWT:287NEWT:246197NEWT:145458NEWT:633149NEWT:1149786NEWT:10239NEWT:68895NEWT:44685NEWT:161934NEWT:4897NEWT:1148NEWT:5744NEWT:72901NEWT:213863NEWT:5508NEWT:3329NEWT:5507NEWT:410661NEWT:55571NEWT:35500NEWT:1235816NEWT:1140NCBITaxon:2157NEWT:1143NEWT:4896NEWT:1287689NEWT:1390NEWT:11557NEWT:1094343NEWT:1462472NEWT:1336795NEWT:644042NEWT:294NEWT:1773NEWT:1182590NEWT:3712NEWT:82380NEWT:3711NEWT:935293NEWT:375146NEWT:270643NEWT:2065263NEWT:177437NEWT:1772NEWT:10418NEWT:118698NEWT:1616117NEWT:263NEWT:118696NEWT:34865NEWT:52283NEWT:11988NEWT:284812NEWT:8175NEWT:400667NEWT:43330NEWT:1603293NEWT:408169NEWT:44664NEWT:47946NEWT:459758NEWT:3702NEWT:1245466NEWT:243277NEWT:1246791NEWT:7067NEWT:118694NEWT:586722NEWT:2850NEWT:118691NEWT:34871NEWT:33548NEWT:274NEWT:7070NCBITaxon:287NEWT:408172NEWT:408170NEWT:96794NEWT:3708NEWT:332648NEWT:44670NEWT:536231NEWT:376219NEWT:219813NEWT:1510NEWT:1436733NEWT:460519NEWT:1247411NEWT:1515NEWT:572307NEWT:1432138NEWT:1424507NEWT:1194599NEWT:272844NEWT:1303443NEWT:9483NEWT:333760NEWT:34607NEWT:56636NEWT:1513458NEWT:1233681NCBITaxon:40559NEWT:506599NEWT:2853422NEWT:81077NEWT:84588NEWT:1679718NEWT:480NEWT:65349NEWT:67767NEWT:46835NEWT:109757NEWT:582580NEWT:300852NEWT:294607NEWT:1502NEWT:128017NEWT:376686NEWT:95486NEWT:508771NEWT:1883446NEWT:253NCBITaxon:2759NEWT:1233435NEWT:93061NEWT:93062NEWT:34613NCBITaxon:4932NEWT:109760NEWT:29031NEWT:943274NEWT:644223NEWT:235443NEWT:108458NEWT:5936NEWT:272623NEWT:272624NEWT:320637NEWT:886559NEWT:3750NEWT:55521NEWT:983964NEWT:118499NEWT:11706NEWT:32644NEWT:527796NEWT:1130798NEWT:55529NEWT:499175NEWT:109779NEWT:3745NEWT:1715989NCBITaxon:4751NEWT:476272NEWT:3747NEWT:195051NEWT:3988NEWT:1116234NEWT:561007NEWT:1255228NEWT:649908NEWT:410289NEWT:373153NEWT:3983NEWT:203696NEWT:472NEWT:352472NEWT:97441NEWT:1071661NEWT:1202531NEWT:1202532NEWT:360094NEWT:1408223NEWT:470NEWT:1408224NEWT:41364NEWT:5911NEWT:1313NEWT:1218097NEWT:1080348NCBITaxon:50557NEWT:1314NEWT:39251NEWT:39491NEWT:212142NCBITaxon:5811NEWT:29491NEWT:101841NEWT:153481NEWT:2014887NEWT:33952NEWT:445NEWT:153009NEWT:261756NEWT:571256NEWT:40483NEWT:63366NEWT:63367NEWT:688333NEWT:215402NCBITaxon:548681NEWT:318586NCBITaxon:5820NEWT:1547NEWT:272634NEWT:198107NEWT:9031NEWT:27292NEWT:1872122NEWT:1308NEWT:7091NEWT:1034015NEWT:760568NEWT:141262NEWT:108931NEWT:1293497NEWT:1055524NEWT:4087NEWT:9778NEWT:306NEWT:150475NEWT:303NEWT:267872NEWT:97477NEWT:7111NEWT:347515NEWT:172269NEWT:9534NEWT:6269NEWT:5180NEWT:256737NEWT:4090NEWT:4092NEWT:9541NEWT:8694NEWT:4093NEWT:33936NEWT:4096NEWT:8692NEWT:2903NEWT:185579NEWT:941442NEWT:536088NEWT:13076NEWT:1226408NEWT:7108NEWT:40479NEWT:43989NEWT:4076NEWT:142615NEWT:317NEWT:1006581NEWT:33940NEWT:885318NEWT:39488NEWT:550NEWT:4081NEWT:273068NEWT:554NEWT:98334NEWT:451516NEWT:552NEWT:231663NEWT:4084NEWT:1325291NEWT:63577NEWT:36185NEWT:1225786NEWT:7574NEWT:1715256NEWT:30640NEWT:575412NEWT:929793NEWT:1507179NEWT:29204NEWT:28112NEWT:114155NEWT:6494NEWT:6491NEWT:507601NEWT:520NEWT:186441NEWT:634955NEWT:643680NEWT:214092NCBITaxon:6157NEWT:6239NEWT:13095NEWT:162425NEWT:104105NEWT:216257NEWT:102169NEWT:208893NEWT:9986NEWT:4054NEWT:8654NEWT:8658NEWT:1268063NEWT:8657NEWT:8655NEWT:5147NEWT:98351NEWT:1263854NEWT:118503NEWT:2059687NEWT:160488NEWT:28104NEWT:207559NEWT:407821NCBITaxon:2NEWT:85057NEWT:985076NEWT:568708NEWT:1215323NEWT:986NEWT:52641NEWT:7159NEWT:267671NEWT:28532NEWT:7160NEWT:353152NEWT:8496NEWT:7165NEWT:1257118NEWT:27448NEWT:40674NEWT:1194669NEWT:51329NEWT:235612NEWT:243230NEWT:443906NEWT:1080772NEWT:519NEWT:8479NEWT:8485NEWT:6063NEWT:49240NEWT:1328388NEWT:104395NEWT:1548728NEWT:667127NEWT:9557NEWT:9315NEWT:6289NEWT:377586NEWT:6287NEWT:38569NEWT:300641NEWT:727NEWT:39655NEWT:9796NEWT:9554NEWT:118562NEWT:725NEWT:7137NEWT:38323NEWT:469008NEWT:256318NEWT:9321NEWT:206411NCBITaxon:6191NEWT:596153NEWT:229533NEWT:1836NEWT:214053NCBITaxon:10298NEWT:80863NEWT:90675NEWT:105231NEWT:104385NEWT:74649NEWT:52638NEWT:57075NEWT:206672NEWT:4097NEWT:8697NEWT:8695NEWT:884204NEWT:71139NEWT:9785NEWT:6279NEWT:1123869NEWT:9544NEWT:9545NEWT:979NEWT:7370NEWT:9793NEWT:83906NCBITaxon:780NCBITaxon:1502NEWT:6282NEWT:1134506NEWT:255470NEWT:27457NEWT:38783NEWT:4006NEWT:6426NCBITaxon:5476NEWT:113208NEWT:33090NEWT:6669NEWT:9935NEWT:89453NEWT:287889NEWT:1114301NEWT:305959NEWT:51515NEWT:13249NEWT:588596NEWT:51511NEWT:8845NEWT:92867NEWT:92866NEWT:419610NEWT:5334NEWT:51750NEWT:202950NEWT:382352NEWT:413071NCBITaxon:10239NCBITaxon:714NEWT:295027NCBITaxon:11320NEWT:632957NEWT:11051NEWT:9925NEWT:89462NCBITaxon:9606NEWT:452680NEWT:8839NEWT:90690NEWT:1436183NEWT:4232NEWT:2758385NEWT:416348NEWT:11298NEWT:171101NEWT:714NEWT:5322NEWT:421932NEWT:312017NEWT:196627NEWT:576131NEWT:200308NEWT:553470NEWT:200302NEWT:870435NEWT:284593NEWT:100392NEWT:655819NEWT:5315NEWT:11082NEWT:9913NEWT:265872NEWT:9915NEWT:105841NEWT:337330NEWT:2666255NEWT:2285NEWT:2287NEWT:6645NEWT:999810NCBITaxon:5693NEWT:28995NEWT:1392998NEWT:380394NEWT:981087NEWT:180066NEWT:226900NEWT:1266738NEWT:231490NEWT:244704NEWT:7725NCBITaxon:6312NEWT:11072NEWT:430615NEWT:563041NEWT:72664NEWT:174633NEWT:326423NEWT:452467NEWT:198822NEWT:36111NEWT:326424NEWT:1416333NEWT:35024NEWT:135461NEWT:1678078NEWT:7962NEWT:749906NEWT:418985NEWT:1817808NEWT:749907NEWT:150847NEWT:145262NEWT:145263NEWT:431947NEWT:69014NEWT:142809NEWT:694569NEWT:130821NEWT:27606NEWT:9739NEWT:1519788NEWT:59202NEWT:9975NEWT:8643NEWT:51953NEWT:502780NEWT:57267NEWT:3197NEWT:13686NEWT:2750454NEWT:860688NEWT:13443NEWT:8644NEWT:1402135NEWT:36329NEWT:62615NEWT:5141NEWT:46360NCBITaxon:3044782NEWT:202914NEWT:1333550NEWT:160235NEWT:502779NEWT:297246NEWT:72407NEWT:8639NEWT:1928434NEWT:8637NEWT:349741NEWT:1114970NEWT:409859NEWT:157295NEWT:2096NEWT:1263871NEWT:509169NEWT:192222NEWT:9721NEWT:137221NEWT:9963NEWT:1617910NEWT:565050NEWT:409860NEWT:2785785NEWT:411901NEWT:8626NEWT:105884NCBITaxon:9615NEWT:8621NEWT:58334NEWT:452659NEWT:1193501NEWT:58331NEWT:6689NEWT:8624NEWT:5116NEWT:8623NEWT:5597NCBITaxon:3077NCBITaxon:12637NEWT:298315NEWT:46105NEWT:8618NEWT:70448NEWT:9704NEWT:1049056NEWT:698936NEWT:1009874NEWT:4498NEWT:8855NEWT:9940NEWT:3168NEWT:10185NEWT:1247190NEWT:521001NEWT:192011NEWT:1813887NEWT:118702NEWT:118700NEWT:2126719NEWT:224911NEWT:118705NCBITaxon:11552NEWT:334249NEWT:6945NEWT:184922NEWT:6703NEWT:2104NEWT:6943NEWT:2NEWT:157546NEWT:35670NEWT:511693NEWT:870726NEWT:4528NEWT:91374NEWT:91373NEWT:474186NEWT:10136NEWT:38942NEWT:35677NEWT:1493NEWT:307972NEWT:1496NEWT:2102NEWT:3674NEWT:660469NEWT:935568NEWT:34NEWT:1365186NEWT:38949NEWT:45474NEWT:1299114NEWT:1590841NEWT:2282334NEWT:4513NEWT:1247NEWT:70667NEWT:869728NEWT:5843NEWT:6938NCBITaxon:39054NCBITaxon:10407NEWT:1736309NEWT:4751NEWT:10360NEWT:194963NEWT:47664NEWT:260799NEWT:317548NEWT:15819NEWT:3654NEWT:237561NEWT:1085956NEWT:5833NEWT:3659NEWT:6928NEWT:1239NEWT:59620NEWT:36745NEWT:499229NEWT:31298NEWT:400727NEWT:1204162NEWT:272563NEWT:11009NEWT:82688NEWT:5823NEWT:3885NEWT:3888NEWT:5821NEWT:44139NEWT:649408NCBITaxon:4895NCBITaxon:4896NEWT:315393NEWT:6915NEWT:3649NEWT:101510NEWT:28903NEWT:325176NEWT:3880NEWT:272559NEWT:216816NEWT:153151NEWT:28909NEWT:515849NEWT:3641NEWT:10141NEWT:95767NEWT:466585NEWT:1000589NEWT:85963NEWT:29NEWT:85962NEWT:42197NEWT:143361NCBITaxon:5796NEWT:12993NEWT:398580NEWT:4565NEWT:1264690NEWT:515619NEWT:1299NEWT:424718NEWT:316407NEWT:34305NEWT:1794699NCBITaxon:183674NEWT:83558NEWT:167387NEWT:84645NEWT:44586NEWT:1123294NEWT:626528NEWT:139927NEWT:4558NEWT:209285NEWT:5888NEWT:211586NEWT:747078NEWT:1282NEWT:1283NEWT:431241NEWT:4550NEWT:1000561NEWT:2133NEWT:4555NEWT:294381NEWT:197NEWT:1390363NEWT:35889NEWT:406072NEWT:1454958NEWT:144030NEWT:288705NEWT:29176NEWT:392815NEWT:4787NEWT:751585NEWT:3217NEWT:113549NEWT:4547NEWT:3218NEWT:180788NEWT:556182NEWT:1270NEWT:374990NEWT:1432059NEWT:154293NEWT:388919NEWT:156471NEWT:4784NEWT:3694NEWT:360106NEWT:156476NEWT:4543NEWT:4785NEWT:210007NEWT:360104NEWT:1857017NEWT:10117NEWT:10359NEWT:10116NEWT:1280NEWT:10358NEWT:83333NEWT:1735272NEWT:83334NEWT:109298NEWT:83332NEWT:317513NEWT:580240NEWT:294128NEWT:11676NEWT:100226NEWT:718308NEWT:1263NEWT:4530NEWT:176299NEWT:5861NEWT:54126NEWT:75058NEWT:13616NEWT:296543NEWT:173NEWT:316435NEWT:42157NEWT:1895NEWT:1034304NEWT:105023NEWT:866628NEWT:64152NEWT:4924NEWT:749200NEWT:2748NEWT:8049NEWT:33892NEWT:9378NEWT:990346NEWT:145953NEWT:257309NEWT:100816NEWT:203124NEWT:192952NEWT:1640346NEWT:230741NEWT:8296NEWT:117008NCBITaxon:1313NEWT:167748NEWT:1163394NEWT:44544NEWT:529818NEWT:400540NEWT:4911NEWT:645463NEWT:152480NEWT:129249NEWT:1077286NEWT:943129NEWT:1221594NEWT:990119NEWT:77216NEWT:1524260NEWT:1111708NEWT:227321NEWT:493760NEWT:93163NEWT:281677NEWT:106590NEWT:260710NEWT:257313NEWT:3827NEWT:400772NEWT:128161NEWT:930945NEWT:1162296NEWT:106592NEWT:106350NEWT:46704NEWT:482428NEWT:1187947NEWT:269796NEWT:10312NCBITaxon:1773NEWT:54571NEWT:190304NEWT:412754NEWT:9598NEWT:8030NEWT:1423772NEWT:53232NEWT:1235987NEWT:1639NEWT:188229NEWT:246437NEWT:3818NEWT:4909NEWT:759272NEWT:432359NEWT:1182263NEWT:943146NEWT:2711NEWT:537457NEWT:479249NEWT:9103NEWT:715877NEWT:862238NEWT:29159NEWT:1155748NEWT:10306NEWT:9595NEWT:8022NEWT:6087NEWT:145943NEWT:595536NEWT:240906NEWT:593117NEWT:89920NEWT:29158NEWT:3633NEWT:319761NEWT:3635NEWT:5811NEWT:3879NEWT:180923NEWT:1328881NEWT:411483NEWT:76336NEWT:3870NEWT:884019NEWT:198215NEWT:411490NEWT:200298NEWT:169963NEWT:1400870NEWT:587NEWT:588NEWT:585NEWT:343NCBITaxon:43925NEWT:84005NEWT:225117NEWT:746128NEWT:8090NEWT:5801NEWT:43665NEWT:212035NEWT:5802NCBITaxon:1653394NEWT:1266464NEWT:1205NEWT:214693NEWT:367830NEWT:178616NEWT:95145NEWT:358NEWT:375451NEWT:357NEWT:9172NEWT:2770979NEWT:2010198NEWT:1277351NEWT:1594786NEWT:457428NEWT:106178NEWT:411469NEWT:84023NEWT:1679NEWT:559292NEWT:214684NEWT:569NEWT:411464NEWT:411460NEWT:2762NEWT:1174673NEWT:1328426NEWT:562NEWT:28035NEWT:411470NEWT:1313166NEWT:2094720NCBITaxon:2697049NCBITaxon:138NEWT:28038NEWT:294750NEWT:1663NEWT:62141NEWT:1422NEWT:1423NEWT:4932NEWT:3603NEWT:2756NEWT:2759NEWT:3848NEWT:3847NEWT:38293NEWT:1428NEWT:198466NEWT:339NEWT:327159NEWT:178876NEWT:1660NEWT:2751NEWT:327160NEWT:573NEWT:571NEWT:142479NEWT:12922NEWT:163159NEWT:29595NEWT:1306413NEWT:241368NEWT:2813599NEWT:518766NEWT:42528NEWT:2664373NEWT:190802NEWT:1213463NEWT:9417NEWT:1216979NEWT:7237NEWT:264483NEWT:509924NEWT:28450NEWT:115104NEWT:1121114NEWT:5061NEWT:663NEWT:8570NEWT:1081927NEWT:1238993NEWT:1408434NEWT:67825NEWT:187878NEWT:1241935NEWT:220668NEWT:1821314NEWT:1249668NEWT:426430NEWT:9407NEWT:37181NEWT:9646NEWT:5286NEWT:7227NEWT:7469NEWT:9402NEWT:9644NEWT:415540NEWT:336722NEWT:675060NEWT:416870NEWT:334542NEWT:313625NEWT:27592NEWT:5052NEWT:426428NEWT:227289NEWT:63459NEWT:2652157NEWT:2652158NEWT:1276815NEWT:40324NEWT:588858NEWT:9638NEWT:9639NCBITaxon:11071NEWT:242231NEWT:197221NEWT:7215NEWT:51453NEWT:2172103NCBITaxon:59201NEWT:7460NEWT:504457NEWT:434271NEWT:392227NEWT:747679NEWT:66712NEWT:42752NEWT:103372NEWT:746360NEWT:862751NEWT:89764NEWT:470150NEWT:9627NEWT:543734NEWT:1009714NEWT:73239NEWT:38018NEWT:226186NEWT:9870NEWT:8782NEWT:30538NEWT:115357NEWT:435590NEWT:1902NEWT:63436NEWT:1908NEWT:1169740NEWT:13164NEWT:1601067NEWT:216129NEWT:1245012NEWT:2664179NEWT:623NEWT:7038NEWT:6192NCBITaxon:1639NCBITaxon:38727NEWT:74790NEWT:2829NEWT:366581NEWT:757417NEWT:216599NEWT:405534NEWT:216595NEWT:40559NEWT:51208NEWT:216597NEWT:9689NEWT:8355NEWT:633NEWT:1607839NEWT:9685NEWT:7029NEWT:6182NEWT:1283300NEWT:9694NEWT:1218707NEWT:186990NEWT:8364NEWT:632NEWT:1204551NEWT:6183NEWT:99586NEWT:630NCBITaxon:11019NEWT:334747NEWT:3907NEWT:61235NEWT:392021NEWT:343691NEWT:1457551NEWT:15368NEWT:1491466NEWT:436486NEWT:340188NEWT:51655NEWT:1933298NEWT:170187NEWT:260707NEWT:121845NEWT:8595NEWT:7263NEWT:186763NEWT:185431NEWT:216591NEWT:29760NEWT:260704NEWT:1719NEWT:703612NEWT:260705NEWT:1202347NEWT:692672NEWT:2697049NEWT:1505907NEWT:1216981NEWT:1109443NEWT:1249885NEWT:7004NEWT:1972855NEWT:9669NCBITaxon:6073NEWT:1089448NEWT:1415167NEWT:1329349NEWT:607699NEWT:6161NEWT:208963NEWT:208964NEWT:27337NEWT:575584NEWT:181088NEWT:185687NEWT:3039NEWT:669202NEWT:2706014NEWT:8727NEWT:8726NEWT:6306NEWT:10090NEWT:8729NEWT:4120NEWT:8720NEWT:5693NEWT:2186NEWT:8724NEWT:8723NEWT:283166NEWT:37334NEWT:5697NEWT:6544NCBITaxon:10357NCBITaxon:10114NCBITaxon:10359NEWT:1685931NEWT:1203258NEWT:242619NEWT:49317NEWT:557723NEWT:557722NEWT:160791NEWT:1353008NEWT:34073NEWT:696811NEWT:402042NEWT:9808NEWT:373995NEWT:5689NEWT:8715NEWT:839NEWT:544404NEWT:92519NEWT:2173NEWT:943925NEWT:4111NEWT:37349NEWT:4113NEWT:837NEWT:640459NEWT:72568NEWT:36015NEWT:51641NEWT:1232801NEWT:81866NEWT:1262469NEWT:5691NEWT:48479NEWT:61435NEWT:627025NEWT:1097677NEWT:375939NEWT:61674NEWT:2072580NEWT:669465NEWT:3258NEWT:1117957NEWT:6526NEWT:112018NEWT:8709NEWT:4100NEWT:1159556NEWT:5671NEWT:1076NEWT:4102NEWT:4101NEWT:6763NEWT:8700NEWT:803NEWT:29722NEWT:1692259NEWT:1486917NEWT:419947NEWT:2847306NEWT:468911NEWT:13373NEWT:817NEWT:7604NEWT:6756NEWT:818NEWT:135588NEWT:1843183NEWT:5661NEWT:157592NEWT:1061NEWT:51866NEWT:29719NEWT:58002NEWT:13136NEWT:5665NEWT:6754NEWT:1179777NEWT:6997NEWT:452589NEWT:509068NEWT:5666NEWT:4577NEWT:813NEWT:5664NEWT:2157NEWT:146479NEWT:1042986NEWT:1911079NEWT:69373NEWT:1843192NEWT:206189NEWT:427577NEWT:264203NEWT:1480154NEWT:9859NEWT:1274414NEWT:9612NEWT:38626NEWT:1286640NEWT:38865NEWT:3077NEWT:9615NEWT:10299NEWT:6100NEWT:713604NEWT:112273NEWT:1087439NEWT:33178NEWT:1147787NEWT:61408NEWT:1440772NEWT:1355477NEWT:8759NEWT:9605NEWT:9606NEWT:90371NEWT:641501NEWT:4153NEWT:9601NEWT:278839NEWT:35128NEWT:39966NEWT:7668NEWT:8756NEWT:915099NEWT:74940NEWT:13551NEWT:10041NEWT:9600NEWT:1450511NEWT:1143193NEWT:1122233NEWT:890382NEWT:174934NEWT:179392NEWT:2823525NEWT:115778NEWT:59540NEWT:9838NEWT:9839NEWT:546414NEWT:227086NEWT:3055NEWT:11167NEWT:9835NEWT:7650NEWT:4146NEWT:5478NEWT:6326NEWT:7656NEWT:5476NEWT:1450520NEWT:1042311NEWT:1123352NEWT:1882945NEWT:746707NEWT:299767NEWT:235909NEWT:28835NEWT:1206781NEWT:47311NEWT:1274432NEWT:127785NEWT:708248NEWT:2053667NEWT:1274426NEWT:104660NEWT:9825NEWT:1274423NEWT:393765NEWT:652611NEWT:104421NEWT:39946NEWT:9823NEWT:95602NEWT:8732NEWT:314539NEWT:763037NEWT:39947NEWT:718274NEWT:1274420NEWT:578458NEWT:6052042055332