#METABOLOMICS WORKBENCH ychen209_20240416_020443 DATATRACK_ID:4778 STUDY_ID:ST003174 ANALYSIS_ID:AN005210 PROJECT_ID:PR001819 VERSION 1 CREATED_ON April 18, 2024, 8:44 am #PROJECT PR:PROJECT_TITLE O-GlcNAcase activity maintains stress granules for proximity-enhanced ATP PR:PROJECT_TITLE production to ensure recovery from stress PR:PROJECT_SUMMARY Accurate disassembly of stress granules (SGs) after environmental stimuli PR:PROJECT_SUMMARY release is essential for cells to maintain homeostasis , which requires PR:PROJECT_SUMMARY ATP-consuming processes. However, the molecular mechanism whereby regulation of PR:PROJECT_SUMMARY SGs programmatically disassembly and ATP restoration remain poorly understood in PR:PROJECT_SUMMARY mammalian cells. Here we found that defect of OGA in cells leads to aggregates PR:PROJECT_SUMMARY formation, severe autophagy and eventually apoptosis during stress recovery. PR:PROJECT_SUMMARY OGA, which localized in SGs, had no effect on SGs formation but could protect PR:PROJECT_SUMMARY SGs from rapid disassembly during stress recovery stage. Then the SGs localized PR:PROJECT_SUMMARY glycolysis-related enzymes were reserved and concentrated in SGs during stress PR:PROJECT_SUMMARY release for ATP production in a proximity manner, which was vital to guarantee PR:PROJECT_SUMMARY cells resistant to stress and survival during recovery. Finally, supplementation PR:PROJECT_SUMMARY of ATP to OGA knockdown cells during stress recovery significantly rescue cell PR:PROJECT_SUMMARY from aggregates, autophagy and apoptosis. Together, these results describe a PR:PROJECT_SUMMARY brand new mechanism on how OGA regulates the programmed disassembly of stress PR:PROJECT_SUMMARY granules and restoration of ATP to safeguard cell viability in a very precisely PR:PROJECT_SUMMARY programmed process, whose rate is rigorous regulated. PR:INSTITUTE Zhejiang University PR:DEPARTMENT Life Sciences Institute PR:LABORATORY Shixian Lin PR:LAST_NAME Chen PR:FIRST_NAME Yulin PR:ADDRESS Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang province, China PR:EMAIL ychen209@qq.com PR:PHONE 18868107794 #STUDY ST:STUDY_TITLE O-GlcNAcase activity maintains stress granules for proximity-enhanced ATP ST:STUDY_TITLE production to ensure recovery from stress (Part 3) ST:STUDY_SUMMARY Accurate disassembly of stress granules (SGs) after environmental stimuli ST:STUDY_SUMMARY release is essential for cells to maintain homeostasis , which requires ST:STUDY_SUMMARY ATP-consuming processes. However, the molecular mechanism whereby regulation of ST:STUDY_SUMMARY SGs programmatically disassembly and ATP restoration remain poorly understood in ST:STUDY_SUMMARY mammalian cells. Here we found that defect of OGA in cells leads to aggregates ST:STUDY_SUMMARY formation, severe autophagy and eventually apoptosis during stress recovery. ST:STUDY_SUMMARY OGA, which localized in SGs, had no effect on SGs formation but could protect ST:STUDY_SUMMARY SGs from rapid disassembly during stress recovery stage. Then the SGs localized ST:STUDY_SUMMARY glycolysis-related enzymes were reserved and concentrated in SGs during stress ST:STUDY_SUMMARY release for ATP production in a proximity manner, which was vital to guarantee ST:STUDY_SUMMARY cells resistant to stress and survival during recovery. Finally, supplementation ST:STUDY_SUMMARY of ATP to OGA knockdown cells during stress recovery significantly rescue cell ST:STUDY_SUMMARY from aggregates, autophagy and apoptosis. Together, these results describe a ST:STUDY_SUMMARY brand new mechanism on how OGA regulates the programmed disassembly of stress ST:STUDY_SUMMARY granules and restoration of ATP to safeguard cell viability in a very precisely ST:STUDY_SUMMARY programmed process, whose rate is rigorous regulated. This is a continuation of ST:STUDY_SUMMARY study ST002927 and ST002936 where an in vitro reaction was performed with ST:STUDY_SUMMARY 13C-glucose on purified stress granules, to validate the conclusion. ST:INSTITUTE Zhejiang University ST:DEPARTMENT Life Sciences Institute ST:LABORATORY Shixian Lin ST:LAST_NAME Chen ST:FIRST_NAME Yulin ST:ADDRESS Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang province, China ST:EMAIL ychen209@qq.com ST:PHONE 18868107794 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - QC-1 Sample source:cultured cells | Factor:QC RAW_FILE_NAME(Raw file name)=00QC; Replicate=QC-1 SUBJECT_SAMPLE_FACTORS - QC-2 Sample source:cultured cells | Factor:QC RAW_FILE_NAME(Raw file name)=00QC; Replicate=QC-2 SUBJECT_SAMPLE_FACTORS - QC-3 Sample source:cultured cells | Factor:QC RAW_FILE_NAME(Raw file name)=00QC; Replicate=QC-3 SUBJECT_SAMPLE_FACTORS - C1 Sample source:cultured cells | Factor:0hr RAW_FILE_NAME(Raw file name)=01C1; Replicate=0hr-1 SUBJECT_SAMPLE_FACTORS - C2 Sample source:cultured cells | Factor:0hr RAW_FILE_NAME(Raw file name)=02C2; Replicate=0hr-2 SUBJECT_SAMPLE_FACTORS - C3 Sample source:cultured cells | Factor:0hr RAW_FILE_NAME(Raw file name)=03C3; Replicate=0hr-3 SUBJECT_SAMPLE_FACTORS - C4 Sample source:cultured cells | Factor:1hr RAW_FILE_NAME(Raw file name)=04C4; Replicate=1hr-1 SUBJECT_SAMPLE_FACTORS - C5 Sample source:cultured cells | Factor:1hr RAW_FILE_NAME(Raw file name)=05C5; Replicate=1hr-2 SUBJECT_SAMPLE_FACTORS - C6 Sample source:cultured cells | Factor:1hr RAW_FILE_NAME(Raw file name)=06C6; Replicate=1hr-3 SUBJECT_SAMPLE_FACTORS - C7 Sample source:cultured cells | Factor:3hr RAW_FILE_NAME(Raw file name)=07C7; Replicate=3hr-1 SUBJECT_SAMPLE_FACTORS - C8 Sample source:cultured cells | Factor:3hr RAW_FILE_NAME(Raw file name)=08C8; Replicate=3hr-2 SUBJECT_SAMPLE_FACTORS - C9 Sample source:cultured cells | Factor:3hr RAW_FILE_NAME(Raw file name)=09C9; Replicate=3hr-3 #COLLECTION CO:COLLECTION_SUMMARY Care was taken to quench cells quickly to minimize oxidation and degradation. CO:SAMPLE_TYPE in vitro reaction #TREATMENT TR:TREATMENT_SUMMARY Stress granules purified from HeLa cells were treated with 13C-Glucose for 0, 1, TR:TREATMENT_SUMMARY 3 hr reaction. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cells without stress treatment or treated with 0.6 M sorbitol stress for 1.5 h SP:SAMPLEPREP_SUMMARY were scraped off and centrifuged to collect cell pellets. The cell pellets were SP:SAMPLEPREP_SUMMARY thawed on ice, resuspended in stress granule lysis buffer (50 mM Tris, 100 mM SP:SAMPLEPREP_SUMMARY K2OAc, 2 mM MgOAc, 0.5 mM DTT, 50 μg/mL heparin, 0.5% NP-40, 0.02% antifoam B, SP:SAMPLEPREP_SUMMARY proteinase inhibitor cocktail, pH 7.4) and lysis by syringe. Lysates were SP:SAMPLEPREP_SUMMARY centrifuged at 1000 × g for 5 min to collect supernatants. Supernatants were SP:SAMPLEPREP_SUMMARY centrifuge at 18000 × g for 20 min to collect pellets and the pellets were SP:SAMPLEPREP_SUMMARY washed once with stress granule lysis buffer. Then, pellets were resuspended in SP:SAMPLEPREP_SUMMARY stress granule lysis buffer and centrifuged at 850 × g for 2 min to collect SP:SAMPLEPREP_SUMMARY supernatants containing stress granule. Then NAD+, ATP and 13C-glucose were SP:SAMPLEPREP_SUMMARY added for reaction at 37 'C for 0, 1 and 3 hr. The reaction was quenched by SP:SAMPLEPREP_SUMMARY 3-fold methanol and centrifuge to collect the supernant for analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Shimadzu 20AD CH:COLUMN_NAME Merck SeQuant ZIC-HILIC (150 x 2.1mm,5um) CH:SOLVENT_A 100% water; 20 mM ammonium carbonate CH:SOLVENT_B 100% acetonitrile CH:FLOW_GRADIENT 0.01 min 80% B, 20 min 20% B, 20.5 min 80% B, 24 min 80% B. CH:FLOW_RATE 0.15 mL/min CH:COLUMN_TEMPERATURE 45 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex Triple Quad 5500+ MS:INSTRUMENT_TYPE QTRAP MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS The raw data were extracted with the software Analyst v1.7.2 and OS v1.7. This MS:MS_COMMENTS is a continuation of study ST002927 where a different set precursor/product ions MS:MS_COMMENTS were chosen for MS. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak intensity MS_METABOLITE_DATA_START Samples QC-1 QC-2 QC-3 C1 C2 C3 C4 C5 C6 C7 C8 C9 Factors Sample source:cultured cells | Factor:QC Sample source:cultured cells | Factor:QC Sample source:cultured cells | Factor:QC Sample source:cultured cells | Factor:0hr Sample source:cultured cells | Factor:0hr Sample source:cultured cells | Factor:0hr Sample source:cultured cells | Factor:1hr Sample source:cultured cells | Factor:1hr Sample source:cultured cells | Factor:1hr Sample source:cultured cells | Factor:3hr Sample source:cultured cells | Factor:3hr Sample source:cultured cells | Factor:3hr CDP 52777.18678 41455.99358 57427.78379 15655.97063 24523.68069 19192.59217 56098.82271 46517.1761 51048.11451 64694.84401 64226.52374 55874.38199 ADP 23019778.86 22654658.25 28817890.58 13967619.65 15432293.77 16855522.34 40343282.05 30902116.3 36275196.72 23022488.26 25707681.11 27193040.21 UDP 8192.006232 5061.707324 16180.00379 2504.188904 1252.011771 2793.046312 10440.14155 11076.31786 9364.914293 13975.6331 25716.14449 19455.42706 GDP 963.2062765 2022.622333 962.2690669 N/A 866.6300875 577.8436865 2413.044039 N/A 1154.461876 577.57372 N/A N/A CTP 22345.50836 16272.01729 22633.49341 674.2026481 577.8660006 577.9339862 21853.21362 17490.80725 20134.7334 46194.29495 47680.67034 38140.46075 UTP 3949.176026 3558.26406 481.5723255 673.8484403 N/A 577.933918 N/A N/A 481.6116277 N/A 866.8419766 N/A ATP 84931666.69 79644155.82 98604051.85 141918593.8 157708470 157987367.1 78668704.54 60877040.4 86761424.63 5995477.017 7200313.723 8069449.839 GTP 1348.492906 2118.933966 N/A 288.9669772 385.2893022 1155.848262 385.2892952 N/A N/A N/A 288.9669772 385.2213429 NAD+ 31593305.32 33474552.6 39499659.38 40195333.4 31494320.61 43361434.08 32915390.49 34407287.55 35279055.59 32964073.26 35408268.24 45997772.65 NADH 89587.13497 87945.21385 98151.64123 62901.73813 55087.83994 88041.7272 149223.084 93242.96758 99406.49561 49934.76768 119057.461 90063.05225 NADP+ 4623.209937 3183.600955 5104.541736 4912.098191 1300.203326 3766.659959 4194.87673 7223.733897 2167.138081 4815.764236 3173.233034 5008.321704 NADPH 288.9669766 N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A pyruvate 45318.6793 280091.2175 47187.5095 52180.79654 9259.467496 34022.54301 38744.4788 29818.57407 6041.34783 45061.3809 50398.7078 84666.25823 pyruvate_C(M+1) 36433.90908 78663.8045 3938.545553 29336.84156 6246.472939 19490.35182 25565.47233 41406.33855 26442.56319 15298.15055 21524.78945 13824.36433 pyruvate_C(M+2) 443579.1102 430467.9665 480006.6561 149787.3848 227333.3033 191039.5079 126456.2204 109507.1787 180571.4048 126647.6527 151133.8317 331655.2448 pyruvate_C(M+3) 9636.39772 9246.229262 12713.18875 2985.752679 4719.184477 5586.122219 3081.745014 2503.872514 5586.278677 5102.084359 4718.198939 7606.883633 glucose 42841.26107 36726.57334 46857.15925 35634.62773 34168.14052 58721.30315 40787.36979 55372.62659 41410.2102 35241.82126 39785.30641 40261.58091 glucose_C(M+1)1 2695.452784 N/A 3370.423124 N/A 1636.186236 1637.327681 2118.145399 1642.506325 1733.689322 1925.657838 2695.799924 2504.180329 glucose_C(M+1)2 1829.191253 N/A 1632.120309 1443.951461 1632.245678 N/A 2310.982449 4623.141964 1637.30062 1444.109395 1540.468098 2600.428326 glucose_C(M+2)1 2078239.373 2102881.449 2228262.244 1963981.025 1961605.345 2329176.724 2107405.4 2005396.408 2239592.671 2051719.938 1987550.36 2196224.668 glucose_C(M+2)2 3226.592016 4045.213326 4141.47562 3659.952687 5201.004306 6549.490305 4719.256079 5489.790079 5393.664311 5200.993669 4719.630577 6164.223288 glucose_C(M+2)3 32845.35986 33030.49005 37561.74466 28796.68192 29084.63053 32162.03035 33222.72135 28752.15666 29402.54478 37561.81237 29856.21166 34190.57033 glucose_C(M+3)1 75577.6541 68683.86176 89863.64736 92369.08373 68774.19105 77110.15402 76606.46117 75413.44135 81957.21431 75899.25658 67375.92951 69715.69679 glucose_C(M+3)2 92763.36056 99302.23741 80612.30098 69183.95391 75755.35649 82306.30646 75852.13348 93811.73003 81525.88542 74138.28382 79177.38198 91201.77275 glucose_C(M+3)3 18010.77571 10199.09777 12280.22405 10546.39202 9347.744513 17625.65724 12477.8158 7973.352579 13816.10034 11826.07274 12971.40547 10108.10971 glucose_C(M+3)4 2574.67401 7319.830421 4521.665308 3365.949326 6453.063326 7319.860712 7127.336634 2889.427641 8475.731338 7030.833739 5971.59562 7608.833004 glucose_C(M+4)1 493812.4061 449859.9422 477742.8417 394583.0763 522421.0837 579387.5747 479110.6173 373631.3177 516069.8935 481477.4413 477814.1709 469836.487 glucose_C(M+4)2 219203.9951 208346.3835 229594.0437 189244.425 192651.8985 257635.9383 238140.0472 199608.3883 386656.5719 230516.4167 196434.3662 229621.5648 glucose_C(M+4)3 160140.5484 194781.5627 178588.5965 148482.4436 158863.3195 180560.2432 168448.6766 157532.7788 164206.9069 180713.8756 148347.2483 280682.5591 glucose_C(M+5)1 2943995.216 2963412.025 3309658.709 2785395.961 3183666.059 3209865.376 3102004.102 2941390.005 3174529.554 2855281.92 2955753.847 3219926.252 glucose_C(M+5)2 5197859.37 4912053.377 4890466.744 4092825.042 3850661.004 5035930.415 5179660.908 7318801.5 4433682.822 4788795.351 3982222.994 4725309.922 glucose_C(M+6) 82469430.55 83332999.03 95354806.09 73589755.53 87292365.33 88899530.56 82168049.83 78259334.79 88083886.67 78597678.34 80576134.87 87431828.18 glucose-6-phosphate 9094.497594 1637.572691 8169.103475 12380.27225 N/A 11352.31112 17319.33787 660.080465 7058.839871 5919.803635 10967.93238 5675.105978 glucose-6-phosphate_C(M+1) 2021.280646 2497.771912 2022.580356 2984.845548 N/A 962.8694149 1155.382227 1054.353655 1926.311641 N/A 1829.957913 963.1383454 glucose-6-phosphate_C(M+2) 4515.201657 2792.670356 2311.543688 2983.941424 2503.872408 3370.96235 2022.768899 2504.346483 2311.541007 1733.576695 3274.716977 2793.203657 glucose-6-phosphate_C(M+3) 2600.609975 3371.03724 2022.644638 2793.186641 2215.27496 2407.874994 1733.714323 1348.012432 2696.847333 2407.882989 1636.434483 2215.252649 glucose-6-phosphate_C(M+4) 12328.3572 11268.51948 10787.17863 14928.9086 14254.22806 12232.01197 13002.32505 10787.2287 10690.87865 13098.4801 13002.43129 11076.03576 glucose-6-phosphate_C(M+5) 577.9339928 577.8999981 N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A glucose-6-phosphate_C(M+6) 4045.19471 2599.29177 3080.861023 3948.869564 1926.094124 2118.946245 4140.930014 1733.632977 3852.502712 3563.636626 3472.328991 2792.909346 fructose-6-phosphate 26391.45626 26967.15783 37609.13621 26679.38102 23693.56475 12617.45726 6747.092524 12660.55035 7801.53426 28892.09232 60013.04601 28316.94316 fructose-6-phosphate_C(M+1) 3087.140216 3082.015001 2311.501712 2022.596252 2504.178304 1637.338346 577.8999937 1733.68299 N/A 3178.504596 5682.660536 2985.724637 fructose-6-phosphate_C(M+2) 674.1856649 2600.388393 N/A N/A N/A N/A N/A 385.2106979 N/A N/A N/A N/A fructose-6-phosphate_C(M+3) 1540.993701 N/A 1830.073288 577.8436883 2311.580296 N/A N/A N/A 1059.472332 3274.65595 1252.085638 3948.91332 fructose-6-phosphate_C(M+4) N/A 963.1472865 N/A 385.2186817 N/A N/A 963.1499868 963.1839664 N/A N/A N/A N/A fructose-6-phosphate_C(M+5) 2889.39265 5681.045825 7512.732603 N/A N/A N/A 6549.46796 5875.121335 7223.314432 9722.460842 5249.044555 6164.148933 fructose-6-phosphate_C(M+6) 70658.46413 76904.50242 77943.59291 17336.89831 17852.67335 24271.29741 95343.00197 84672.67925 107737.5348 111648.6433 119325.742 124378.0202 fructose-1,6-bisphosphate 385.2893022 1155.671396 N/A N/A N/A 288.9669964 N/A N/A N/A N/A N/A N/A fructose-1,6-bisphosphate_C(M+1) 963.1866475 N/A 963.1473321 N/A N/A N/A N/A N/A 577.8633393 1444.685678 1733.706338 1348.394677 fructose-1,6-bisphosphate_C(M+2) 8668.092739 7801.610915 9824.140324 1348.433978 2118.900013 1926.305303 7319.907971 6452.96602 7416.205375 22056.136 17914.64293 18973.77474 fructose-1,6-bisphosphate_C(M+3) N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A fructose-1,6-bisphosphate_C(M+4) 9038.064276 12906.22754 12857.9687 5875.101723 6356.896576 5007.296461 14880.61923 14533.0358 12997.41368 13693.60744 13960.6743 14155.97599 fructose-1,6-bisphosphate_C(M+5) 4430.480288 4237.891983 3076.879338 N/A 2215.317979 3274.763225 5393.693658 4430.471263 6934.824196 5585.924889 6838.560866 4334.008733 fructose-1,6-bisphosphate_C(M+6) 577.9339796 N/A N/A N/A N/A N/A N/A N/A N/A 770.5106649 N/A N/A dihydroxy-acetone-phosphate 3043.811086 888.0248358 5540.393394 12248.72744 1567.426505 2795.239473 1164.757648 2838.132192 1275.677366 4213.472885 2747.261988 2681.769997 dihydroxy-acetone-phosphate_C(M+1) 261992.864 245403.0933 264149.2214 243899.7945 267558.6496 283224.6141 259743.0536 257144.0492 256276.091 268539.9559 267831.1419 254312.3865 dihydroxy-acetone-phosphate_C(M+2) 1151077.098 1104525.573 1260844.453 298682.9795 317505.2056 361929.2841 1100428.079 1025061.133 1089640.381 2324542.237 2216404.626 2002220.242 dihydroxy-acetone-phosphate_C(M+3) 33304.42944 76071.0668 67746.43368 9539.110579 9263.488799 11460.77273 69220.59785 41296.82654 48995.13804 144023.5912 145126.9894 132429.3299 D-glyceraldehdye-3-phosphate 36556.68883 53649.94387 48937.30701 21178.16893 34789.30918 18161.55331 134068.8097 86345.15732 13093.56157 31233.88322 43239.02612 63854.99513 D-glyceraldehdye-3-phosphate_C(M+1) 2407.938244 1252.116978 674.188289 N/A 1251.880828 N/A 3948.891578 3173.288199 N/A 3365.948226 2499.071148 2980.588041 D-glyceraldehdye-3-phosphate_C(M+2) 33847.1191 67439.47914 60911.39133 60293.87678 102913.8672 48718.86939 58076.61057 66361.56232 51249.79174 69154.14834 26457.95144 41213.75605 D-glyceraldehdye-3-phosphate_C(M+3) 22638.30134 24174.92122 28050.85298 769.8488428 N/A N/A 8340.014955 8994.795382 11262.39176 63147.5233 72739.91579 69109.98895 1,3-diphosphateglycerate 27260.81755 38627.2798 32703.83702 9684.248554 12038.04017 12225.76411 35668.66527 34421.44882 40013.51175 43021.98208 39629.31703 40393.01218 1,3-diphosphateglycerate_C(M+1) 385.2383393 N/A 481.5356913 N/A N/A N/A N/A N/A N/A N/A N/A N/A 1,3-diphosphateglycerate_C(M+2) 674.2392956 N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 1,3-diphosphateglycerate_C(M+3) 866.7856979 1348.299077 577.875 1348.355362 963.1276572 1155.791996 N/A N/A N/A 1348.276751 577.8580102 866.8643153 3-phosphoglycerate 7290.327543 10266.21975 11207.38231 10042.9995 4257.291226 5062.919311 10069.91691 8624.261526 13065.60253 17295.76947 11629.89648 15361.86956 3-phosphoglycerate_C(M+1) 4623.171307 3376.165984 2504.126705 1449.851629 2889.483993 770.5153801 764.7080788 1733.194778 3082.104305 1059.458139 2215.269677 2119.006299 3-phosphoglycerate_C(M+2) 1347.966444 1352.862305 1829.942039 1251.851713 2118.151732 914.966027 12327.87915 1925.756174 3274.680296 2162.001941 2792.692633 4429.70158 3-phosphoglycerate_C(M+3) 3080.521621 3371.024964 3753.492287 N/A N/A N/A 3371.005342 3082.106338 3274.717023 5902.316229 14350.4248 14446.78663 2-phosphoglycerate 23123.64432 24633.26655 38274.98842 11868.45447 33560.82904 14716.3042 38045.62213 43102.28551 9381.543667 17975.67291 19367.69843 10827.95181 2-phosphoglycerate_C(M+1) 17690.73751 21717.13094 10680.12241 4998.118484 13732.40732 4910.505929 12517.36265 13097.55736 16264.36579 4724.60736 11734.37292 10106.7172 2-phosphoglycerate_C(M+2) 4232.172923 5642.319636 5272.039788 2869.931839 8847.585597 2139.15019 5324.835984 4287.720174 2645.65281 2555.660142 7107.537428 6088.939725 2-phosphoglycerate_C(M+3) 10011.64988 3119.9614 14638.80051 N/A 385.2303495 N/A 10584.82217 7699.915628 10396.83032 23493.53803 46580.22549 41987.9903 phosphoenolpyruvate 5682.74657 572.5482633 3081.474697 1444.144567 3852.582954 N/A N/A 1540.484027 2886.938369 4333.012045 N/A 2301.303828 phosphoenolpyruvate_C(M+1) 1347.880218 2407.835678 N/A 963.1643351 2118.863372 N/A 1637.344642 1444.764254 1637.011157 1059.508993 N/A 866.9264 phosphoenolpyruvate_C(M+2) 1155.82867 N/A 385.2893285 1155.867972 N/A N/A 577.8973316 1252.170653 866.9009826 481.5919849 674.1606913 N/A phosphoenolpyruvate_C(M+3) 30626.34092 29952.48003 27159.62756 32263.86583 33999.25864 35542.07841 32265.73286 33711.53642 31011.82547 28797.00257 31780.55327 30241.22499 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Precursor ion (m/z) Product ion (m/z) Retention time (min) PubChem CID CDP 402 384 10.82 6132 ADP 426.1 159 10.34 6022 UDP 403 159 10.24 6031 GDP 442 159 12.06 135398619 CTP 482 384 6176 UTP 483 159 12.6 6133 ATP 506.1 159 11.56 5957 GTP 522 424 13.46 135398633 NAD+ 662 540 7.7 5892 NADH 664 408 7.14 439153 NADP+ 742 620 10.64 5886 NADPH 744 408 10.62 5884 pyruvate 87 43 4.16 107735 pyruvate_C(M+1) 88 44 4.16 pyruvate_C(M+2) 89 45 4.16 pyruvate_C(M+3) 90 45 4.16 glucose 179 88.8 5793 glucose_C(M+1)1 180 88.8 glucose_C(M+1)2 180 89.8 glucose_C(M+2)1 181 88.8 glucose_C(M+2)2 181 89.8 glucose_C(M+2)3 181 90.8 glucose_C(M+3)1 182 88.8 glucose_C(M+3)2 182 89.8 glucose_C(M+3)3 182 90.8 glucose_C(M+3)4 182 91.8 glucose_C(M+4)1 183 89.8 glucose_C(M+4)2 183 90.8 glucose_C(M+4)3 183 91.8 glucose_C(M+5)1 184 90.8 glucose_C(M+5)2 184 91.8 glucose_C(M+6) 185 91.8 glucose-6-phosphate 259.02 199 10.25 5958 glucose-6-phosphate_C(M+1) 260.02 200 10.25 glucose-6-phosphate_C(M+2) 261.02 201 10.25 glucose-6-phosphate_C(M+3) 262.02 202 10.25 glucose-6-phosphate_C(M+4) 263.02 203 10.25 glucose-6-phosphate_C(M+5) 264.02 204 10.25 glucose-6-phosphate_C(M+6) 265.02 205 10.25 fructose-6-phosphate 259.03 169 9.7 69507 fructose-6-phosphate_C(M+1) 260.03 170 9.7 fructose-6-phosphate_C(M+2) 261.03 171 9.7 fructose-6-phosphate_C(M+3) 262.03 172 9.7 fructose-6-phosphate_C(M+4) 263.03 173 9.7 fructose-6-phosphate_C(M+5) 264.03 174 9.7 fructose-6-phosphate_C(M+6) 265.03 175 9.7 fructose-1,6-bisphosphate 339 97 10267 fructose-1,6-bisphosphate_C(M+1) 340 97 fructose-1,6-bisphosphate_C(M+2) 341 97 fructose-1,6-bisphosphate_C(M+3) 342 97 fructose-1,6-bisphosphate_C(M+4) 343 97 fructose-1,6-bisphosphate_C(M+5) 344 97 fructose-1,6-bisphosphate_C(M+6) 345 97 dihydroxy-acetone-phosphate 169 79 8.98 668 dihydroxy-acetone-phosphate_C(M+1) 170 79 8.98 dihydroxy-acetone-phosphate_C(M+2) 171 79 8.98 dihydroxy-acetone-phosphate_C(M+3) 172 79 8.98 D-glyceraldehdye-3-phosphate 169.05 97 9.6 729 D-glyceraldehdye-3-phosphate_C(M+1) 170.05 97 9.6 D-glyceraldehdye-3-phosphate_C(M+2) 171.05 97 9.6 D-glyceraldehdye-3-phosphate_C(M+3) 172.05 97 9.6 1,3-diphosphateglycerate 265 79 439191 1,3-diphosphateglycerate_C(M+1) 266 79 1,3-diphosphateglycerate_C(M+2) 267 79 1,3-diphosphateglycerate_C(M+3) 268 79 3-phosphoglycerate 185 97 12.02 724 3-phosphoglycerate_C(M+1) 186 97 12.02 3-phosphoglycerate_C(M+2) 187 97 12.02 3-phosphoglycerate_C(M+3) 188 97 12.02 2-phosphoglycerate 184.8 97 59 2-phosphoglycerate_C(M+1) 185.8 97 2-phosphoglycerate_C(M+2) 186.8 97 2-phosphoglycerate_C(M+3) 187.8 97 phosphoenolpyruvate 167 79.1 11.14 1005 phosphoenolpyruvate_C(M+1) 168 79.1 11.14 phosphoenolpyruvate_C(M+2) 169 79.1 11.14 phosphoenolpyruvate_C(M+3) 170 79.1 11.14 METABOLITES_END #END