#METABOLOMICS WORKBENCH ychen209_20230919_041456 DATATRACK_ID:4325 STUDY_ID:ST002927 ANALYSIS_ID:AN004801 PROJECT_ID:PR001819 VERSION 1 CREATED_ON October 13, 2023, 10:36 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 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. 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 Factor:QC RAW_FILE_NAME=00QC; Other=QC-1 SUBJECT_SAMPLE_FACTORS - QC-2 Factor:QC RAW_FILE_NAME=00QC; Other=QC-2 SUBJECT_SAMPLE_FACTORS - QC-3 Factor:QC RAW_FILE_NAME=00QC; Other=QC-3 SUBJECT_SAMPLE_FACTORS - C1 Factor:noStressHeLa RAW_FILE_NAME=01C1; Other=noStressHeLa-1 SUBJECT_SAMPLE_FACTORS - C2 Factor:noStressHeLa RAW_FILE_NAME=02C2; Other=noStressHeLa-2 SUBJECT_SAMPLE_FACTORS - C3 Factor:noStressHeLa RAW_FILE_NAME=03C3; Other=noStressHeLa-3 SUBJECT_SAMPLE_FACTORS - C4 Factor:noStressKDOGA RAW_FILE_NAME=04C4; Other=noStressKDOGA-1 SUBJECT_SAMPLE_FACTORS - C5 Factor:noStressKDOGA RAW_FILE_NAME=05C5; Other=noStressKDOGA-2 SUBJECT_SAMPLE_FACTORS - C6 Factor:noStressKDOGA RAW_FILE_NAME=06C6; Other=noStressKDOGA-3 SUBJECT_SAMPLE_FACTORS - C7 Factor:StressHeLa RAW_FILE_NAME=07C7; Other=StressHeLa-1 SUBJECT_SAMPLE_FACTORS - C8 Factor:StressHeLa RAW_FILE_NAME=08C8; Other=StressHeLa-2 SUBJECT_SAMPLE_FACTORS - C9 Factor:StressHeLa RAW_FILE_NAME=09C9; Other=StressHeLa-3 SUBJECT_SAMPLE_FACTORS - C10 Factor:StressKDOGA RAW_FILE_NAME=10C10; Other=StressKDOGA-1 SUBJECT_SAMPLE_FACTORS - C11 Factor:StressKDOGA RAW_FILE_NAME=11C11; Other=StressKDOGA-2 SUBJECT_SAMPLE_FACTORS - C12 Factor:StressKDOGA RAW_FILE_NAME=12C12; Other=StressKDOGA-3 SUBJECT_SAMPLE_FACTORS - C13 Factor:RecoveryHeLa RAW_FILE_NAME=13C13; Other=RecoveryHeLa-1 SUBJECT_SAMPLE_FACTORS - C14 Factor:RecoveryHeLa RAW_FILE_NAME=14C14; Other=RecoveryHeLa-2 SUBJECT_SAMPLE_FACTORS - C15 Factor:RecoveryHeLa RAW_FILE_NAME=15C15; Other=RecoveryHeLa-3 SUBJECT_SAMPLE_FACTORS - C16 Factor:RecoveryKDOGA RAW_FILE_NAME=16C16; Other=RecoveryKDOGA-1 SUBJECT_SAMPLE_FACTORS - C17 Factor:RecoveryKDOGA RAW_FILE_NAME=17C17; Other=RecoveryKDOGA-2 SUBJECT_SAMPLE_FACTORS - C18 Factor:RecoveryKDOGA RAW_FILE_NAME=18C18; Other=RecoveryKDOGA-3 #COLLECTION CO:COLLECTION_SUMMARY Care was taken to quench cells quickly to minimize oxidation and degradation. CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY HeLa cells of shControl or shOGA were treated with no stress, stress by 1 M TR:TREATMENT_SUMMARY sorbitol for 1 hr, or recovery from stress for 2 hr. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Briefly, equal confluency of cells in 6-well plate with 3 biological replicates SP:SAMPLEPREP_SUMMARY were rinsed by 1 mL HPLC-grade water rapidly and immediately quenched by SP:SAMPLEPREP_SUMMARY directly added by 1 mL liquid nitrogen to the plate. Then, 1 mL ice-cold 90% MC SP:SAMPLEPREP_SUMMARY buffer (MC, 9: 1 MeOH: CHCl3. 90% MC, 9 : 1 MC: water) with 5 µM methionine SP:SAMPLEPREP_SUMMARY sulfone as internal signal was added to each well of plate and the cells were SP:SAMPLEPREP_SUMMARY scraped and transferred to1.5-mL tubes. Extracts were centrifuged at 4 ℃ for 5 SP:SAMPLEPREP_SUMMARY min at 16,000 xg and the supernatants were transferred to new clean 1.5-mL tubes SP:SAMPLEPREP_SUMMARY and equal volume of each sample was pooled for the QC sample. Finally, the SP:SAMPLEPREP_SUMMARY extracts were dried using vacuum concentrator system and store at -80 ℃ until SP:SAMPLEPREP_SUMMARY analysis. For targeted analysis, the dried extracts were resuspended by 100 µL SP:SAMPLEPREP_SUMMARY 60% acetonitrile and centrifuged at 4 ℃ for 15 min at 15,000 xg after 30 min SP:SAMPLEPREP_SUMMARY incubation at 4 ℃. Only the supernatant was transferred to autosampler vials SP:SAMPLEPREP_SUMMARY for MS 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. #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 C10 C11 C12 C13 C14 C15 C16 C17 C18 Factors Factor:QC Factor:QC Factor:QC Factor:noStressHeLa Factor:noStressHeLa Factor:noStressHeLa Factor:noStressKDOGA Factor:noStressKDOGA Factor:noStressKDOGA Factor:StressHeLa Factor:StressHeLa Factor:StressHeLa Factor:StressKDOGA Factor:StressKDOGA Factor:StressKDOGA Factor:RecoveryHeLa Factor:RecoveryHeLa Factor:RecoveryHeLa Factor:RecoveryKDOGA Factor:RecoveryKDOGA Factor:RecoveryKDOGA glyoxylate 14892.891 1128.051999 262.3976585 32288.05819 12792.69793 11529.8408 20098.50583 6675.919391 2753.893123 N/A 1887.495527 166.6500396 N/A N/A 3662.206835 10169.17932 N/A 777.7501187 N/A N/A 8233.834277 pyruvate 859656.1384 1008802.504 986468.7311 478013.0876 383633.1115 347872.2493 284065.6829 256709.1361 192468.2997 499936.4151 516766.3693 533755.7418 430731.4167 527085.5792 608688.2267 2472415.517 946915.3947 576136.8157 1945109.445 2432779.425 837842.2024 lactate 16617080.29 16144649.89 16571702.47 35320683.2 30084292.35 28977465.65 19440637.63 15551930.98 11163731.88 7933898.637 8729060.835 7999760.958 11253795.21 8941972.295 6781493.879 23934312.95 11830689.87 13662355.38 14104004.24 19433677.27 10424782.55 fumarate 622914.0293 897298.6989 755757.4333 983477.2662 848643.9611 1076461.612 544149.0571 585273.3974 534696.8648 1057240.592 1032799.482 1445980.489 633156.0234 663134.8472 554336.5136 459383.1539 392290.019 565047.0991 369931.0774 156694.3533 147158.7443 succinate 737807.3162 835026.2179 765887.274 566386.5132 487514.0043 505512.6993 482190.34 397916.6488 302217.277 1858787.373 1706622.704 1663319.312 1041787.973 1087686.399 909510.5129 973199.1856 512468.7245 639880.1519 793960.3098 1047852.74 440535.978 oxaloacetate 202740.6685 169188.3982 N/A 550277.6444 26982.42159 359365.2442 105390.0847 232622.9566 68382.70002 N/A 8708.108703 3054.116974 444.4425803 9982.39522 14385.53396 22338.52374 8631.187602 2659.088747 N/A N/A N/A malate 22654785.99 29346925.02 29682833.28 39704389.45 34736253.59 44405577.1 20519306.41 24324024.18 20938954.78 45518155.62 43085177.1 55334878.36 26438572.81 24712985.52 22069534.42 18020143.42 14392363.29 20935722.52 5182921.099 5257567.3 5017525.757 a-ketoglutarate 2499614.052 2745626.323 2935306.716 2772315.521 3230655.175 3474859.44 2411937.973 2389674.54 1988040.237 5787684.304 5346278.962 5973980.534 2638659.487 1773551.422 3028993.941 2441496.494 2156642.25 3048366.667 1311045.646 1362834.259 753757.3492 phosphoenolpyruvate 8521.462834 9531.670115 26015.99536 19972.68598 15624.23881 9165.195918 20495.79117 24308.01224 20660.44504 22463.41901 8389.702186 14269.7756 9722.034881 10068.26511 16280.54451 25043.33197 18477.94384 23829.23858 17114.37186 14215.98873 16781.39692 D-glyceraldehdye-3-phosphate 1303557.269 1498778.572 1540063.794 1611620.409 1442289.299 1748049.281 1304070.362 1303022.845 1135063.658 1450067.128 1284833.935 1407053.948 1529179.79 1360650.287 1216393.206 2075404.238 1936156.169 2814841.302 1538862.646 1467522.993 1391708.129 aconitate 563377.72 725256.0002 827170.2885 1028253.403 600000.9283 1039753.909 579803.9136 851461.0121 966481.4801 464193.0778 532030.2559 577813.9272 287560.4549 280381.4505 161722.7819 1072694.567 525110.4265 943045.8058 394544.0557 513337.5833 216992.4391 citrate-isocitrate 68262.84238 207634.6567 148427.4706 724801.4908 272096.8502 218915.826 260404.9083 462203.3343 137388.0686 211957.9027 160472.0329 227918.504 61553.08826 45360.64865 84922.98901 373570.18 141865.6033 62244.62703 120209.8042 85751.23299 57175.34813 isocitrate 310270.8063 12278.08123 1554.121864 305558.9162 N/A 50802.98043 89694.88106 168416.2034 9843.816121 N/A 13231.9749 719.1760146 N/A 469.6709674 8174.685373 472.3827162 2872.013472 222.2212902 666.6726465 4369.043245 5775.323115 citrate 85685.36467 57772.34761 73919.93554 55598.08323 28496.69708 57952.4462 40647.47439 110326.827 29308.58652 10009.80321 38699.67341 80524.14913 19894.90353 21397.47263 10221.03209 69449.49306 57193.68718 23114.62427 32500.22022 88993.39385 46003.24928 glucose-6-phosphate 206404.473 225558.1808 263643.1781 416747.0147 339895.6007 427366.6647 255406.2112 389959.8145 378964.3708 112732.4234 91193.5638 138404.9646 84375.76696 87660.10802 46021.33313 245619.7551 239830.514 357836.4437 65810.80278 61877.32168 49968.174 fructose-6-phosphate 144400.2669 183104.8004 174611.0121 349917.569 270302.2488 375030.6147 317263.9949 368821.1438 368182.5354 100754.569 95501.28227 61294.11185 63220.91434 69884.54523 76680.51161 207117.9611 136283.48 285395.0518 32163.94059 61076.26198 70862.40753 1_3-diphopshateglycerate 39281.89912 27186.75631 31597.88584 33046.56109 23228.06543 19104.0562 37719.00798 29898.06215 35568.23788 20928.32996 21472.21168 47587.35376 20711.55348 27594.7827 22123.48184 26002.84122 29679.16973 45393.49519 24212.68053 38468.5525 25645.09225 N-acetyl-glucosamine-1-phosphate 117012.1743 123266.0288 122755.2385 50288.39674 51006.88935 16801.35862 223626.0271 193350.3526 202274.9449 31720.3571 48724.32033 38314.63948 379393.9578 293539.2486 241988.5903 43813.93435 33553.29045 59913.9431 89599.87047 96663.302 70593.08186 fructose-1_6-bisphosphate 207171.7253 N/A 48068.24467 N/A N/A 985.5093144 22837.55613 57422.35965 N/A N/A 3432.623219 553.2611573 1909.643762 61438.70408 2430.343191 310.8919315 790.9548583 2882.330532 1984.18405 357.7073567 N/A CDP_neg 5582.728805 31125.20717 22133.65119 22443.88194 14608.9255 9674.618656 12483.01841 2984.397997 19383.43049 12926.46773 14279.85833 22012.75176 13561.06979 27736.41051 10242.54976 87802.91139 35978.76596 58404.11729 33409.37272 16846.55996 51843.44737 UDP_neg 38724.24624 48502.82143 48619.41459 20306.29969 21864.13819 24804.18516 76290.84335 101401.9249 44188.76667 20264.26071 15895.25397 18115.10475 97076.21868 98215.50026 90158.6737 22946.1063 28630.78547 31144.32379 26569.39056 40336.64372 30342.34009 ADP_neg 105651.6534 181591.6507 146712.7704 94010.26715 109425.9428 160825.7256 227313.0723 164439.9251 287447.9468 112204.5193 369017.4532 448791.0503 122494.9597 171238.9014 110137.4096 27830.23997 56047.09616 177465.8719 165627.1737 67372.9724 43231.89603 GDP_neg 12745.82096 16530.35505 11615.38379 17882.45694 8388.284513 7114.30691 7635.52883 5875.447133 13336.88442 1832.314741 11604.11455 8408.116876 9497.762317 3005.696815 5472.609898 12522.86286 2731.563208 9949.175017 14139.33518 7024.00061 4745.450268 FMN 1055.705323 N/A N/A N/A 2444.61079 1191.817953 1669.727373 500.0273965 503.0773285 1058.247782 1108.236522 1333.376745 2278.031909 1722.331958 1830.622583 3611.61265 1163.817366 1268.911963 777.8553225 1000.212676 1832.986182 CTP_neg 21984.64597 28027.12087 22177.16754 11043.95042 35582.91663 8021.591197 31620.18427 16474.98548 12295.93084 6693.061706 5618.034991 5880.012095 3047.068626 7543.358877 20471.84587 6267.643166 42857.19959 958.3946959 10301.84338 30480.96564 21745.96065 UTP_neg 176640.818 238570.2653 61597.26194 205648.489 160093.3591 184298.9388 91000.11025 74433.61193 188225.2545 126979.1628 349271.5545 120777.7902 113735.947 144083.6223 92824.92905 93071.1076 603069.3403 84739.25369 184328.0227 86859.60814 132492.2087 CDP-choline 296475.6542 315660.0231 325389.5854 34615.0112 35227.32682 38426.3363 48672.77128 43491.89051 29621.02323 230752.6354 242394.3823 282453.8359 406265.2556 386345.0622 447576.358 797528.5309 791198.348 1033070.535 417055.8873 488517.8722 438730.5856 ATP_neg 67498.41044 297967.2014 236555.3403 94104.36308 54782.34357 156126.3667 1225223.66 354875.724 427131.8226 852676.9519 462806.9307 741115.6102 262592.5929 381547.3611 226763.1141 315936.9912 207889.5444 208319.174 213500.561 110918.4619 216934.7543 GTP_neg 2408.617208 33825.29549 37162.08587 45059.70591 38490.60823 91748.19731 34574.20538 13718.61849 91979.00546 81022.71945 69620.97403 99260.30234 44338.80529 11852.12594 13881.87561 69818.27455 49804.38116 13523.74584 48210.35068 15863.51926 11481.80197 UDP-N-acetyl-glucosamine 2482319.286 3113221.896 3017737.514 1242851.112 1255651.258 1452987.631 5616991.352 5815845.262 5142335.864 1335113.502 1194855.364 1266787.691 6350817.726 6499511.98 6182165.788 1756252.577 1555367.197 2183496.416 2039523.954 2501221.162 1905489.714 NAD+_neg 4639777.584 5010336.422 5284903.881 10903558 11395295.07 12039532.67 9616295.677 8110462.092 9272908.243 4519403.623 4211134.249 4416688.348 2727026.412 2737738.196 2150831.039 2669562.069 2578212.299 3455462.878 728930.7295 705386.9497 826830.0363 NADH_neg 25505.79262 18785.0953 28781.71878 48459.99478 57862.0458 52213.56037 40589.95261 39283.05708 47883.52074 23058.4066 22780.66489 23724.47712 17779.83315 19057.83406 15223.85568 27558.22211 23669.81138 13375.7975 8890.51321 4497.676042 9667.640668 NADP+_neg 157596.8874 59452.86491 174793.7046 111666.3937 74398.85364 192315.2157 87010.52629 55363.30204 91898.16794 209918.8285 64125.2408 50741.63992 239187.5862 204924.4833 77898.18814 150024.8459 92424.88393 143959.6233 12773.51956 65014.10553 41186.60657 NADPH_neg 22638.97365 25125.61976 29724.92384 29534.49993 6589.981396 29164.35123 12488.58574 24791.81323 19004.46516 36660.76163 26099.50177 29953.77154 15986.06711 26904.72529 25436.49558 36488.87571 30726.11167 30362.65308 15281.34713 16890.74271 6972.975398 coenzyme A_neg 499.9554268 N/A 277.8446643 N/A N/A N/A 666.8233213 N/A N/A N/A 889.0426262 N/A N/A 608.141991 777.8106724 N/A 833.4327065 1055.710579 333.4679724 388.9393681 222.3306575 acetyl-CoA_pos 5547.569166 5842.894514 4734.560731 4216.5367 3444.694025 3222.516016 2111.318047 N/A 1222.366007 6226.226026 8167.657189 7896.139056 8482.265937 9692.845214 2858.505884 10112.53397 3389.129925 2228.251407 6278.93032 3795.788067 1914.009336 succinyl-CoA-methylmalonyl-CoA_neg 277.9073273 N/A 388.8713297 N/A N/A N/A N/A 277.8286967 N/A 222.2213264 N/A N/A N/A N/A 222.3573141 N/A 611.1606715 N/A 222.2160204 499.9820276 N/A MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Precursor ion (m/z) Product ion (m/z) Retention time (min) glyoxylate 73 45 4.16 pyruvate 87 43 3.66 lactate 89 43.2 3.9 fumarate 115 71 9.7 succinate 117 73 9.06 oxaloacetate 131 87 malate 133 115 9.8 a-ketoglutarate 145 101 9.28 phosphoenolpyruvate 167 79 11.14 D-glyceraldehdye-3-phosphate 169.05 97 9.6 aconitate 173.05 85 11.12 citrate-isocitrate 191 111 11.45 isocitrate 191.02 117 citrate 191.05 87 glucose-6-phosphate 259.02 199 10.25 fructose-6-phosphate 259.03 169 9.7 1_3-diphopshateglycerate 265 79 N-acetyl-glucosamine-1-phosphate 300 79 fructose-1_6-bisphosphate 339 97 CDP_neg 402 384 10.82 UDP_neg 403 159 10.24 ADP_neg 426.1 159 10.34 GDP_neg 442 159 12.06 FMN 455 213 CTP_neg 482 384 UTP_neg 483 159 12.6 CDP-choline 487 428 ATP_neg 506.1 159 11.56 GTP_neg 522 424 13.46 UDP-N-acetyl-glucosamine 606 385 8.95 NAD+_neg 662 540 7.7 NADH_neg 664 408 7.14 NADP+_neg 742 620 10.64 NADPH_neg 744 408 10.62 coenzyme A_neg 766 408 8.15 acetyl-CoA_pos 808 408 7.21 succinyl-CoA-methylmalonyl-CoA_neg 866.2 822 METABOLITES_END #END