#METABOLOMICS WORKBENCH guying_20241009_181944 DATATRACK_ID:5277 STUDY_ID:ST003531 ANALYSIS_ID:AN005800 PROJECT_ID:PR002173 VERSION 1 CREATED_ON October 24, 2024, 5:13 pm #PROJECT PR:PROJECT_TITLE Micropeptide hSPAR, a glutamine regulator, suppresses tumor growth via PR:PROJECT_TITLE TRIM21-P27KIP1-mTOR pathway PR:PROJECT_SUMMARY The mammalian target of rapamycin (mTOR) plays pivotal roles in cancer growth PR:PROJECT_SUMMARY control upon amino acid response. Recently, cyclin-dependent kinase (CDK) PR:PROJECT_SUMMARY inhibitor cyclin-dependent kinase inhibitor 1B (CDKN1B or P27KIP1) 1 has been PR:PROJECT_SUMMARY reported as a non-canonical inhibitor to mTOR signaling in mouse embryo PR:PROJECT_SUMMARY fibroblasts (MEFs). However, the mechanisms underlying P27KIP1-mTOR axis are PR:PROJECT_SUMMARY yet-to-be uncovered. Here, we find that micropeptide human small regulatory PR:PROJECT_SUMMARY polypeptide of amino acid response (hSPAR）, through its C-terminus (hSPAR-C), PR:PROJECT_SUMMARY inhibits E3 ligase tripartite motif containing 21 (TRIM21)-mediated P27KIP1 PR:PROJECT_SUMMARY degradation and causes P27KIP1’s cytoplasmic accumulation in breast cancer PR:PROJECT_SUMMARY cells. Interestingly, hSPAR/hSPAR-C also serves as an inhibitor to glutamine PR:PROJECT_SUMMARY transporter SLC38A2 and remarkably decreases the cellular glutamine level PR:PROJECT_SUMMARY specifically in cancer cells. The resulted glutamine deprivation sequentially PR:PROJECT_SUMMARY triggers translocation of cytoplasmic P27KIP1 to lysosomes, where P27KIP1 PR:PROJECT_SUMMARY disrupts Ragulator complex and suppresses mTOR complex 1 (mTORC1) assembly. PR:PROJECT_SUMMARY Administration of hSPAR or hSPAR-C dramatically impedes breast cancer cell PR:PROJECT_SUMMARY proliferation and xenografic tumor growth. Collectively, we define hSPAR as an PR:PROJECT_SUMMARY intrinsic molecule to control cellular glutamine level and a PR:PROJECT_SUMMARY previously-unidentified tumor suppressor by promoting accumulation and PR:PROJECT_SUMMARY lysosomal-localization of P27KIP1 to inhibit mTORC1 assembly. PR:INSTITUTE University of Science and Technology of China PR:LAST_NAME Wang PR:FIRST_NAME Wei PR:ADDRESS Division of Life Sciences and Medicine, 443 Huangshan Road, Hefei city, Anhui PR:ADDRESS Province, 230022, China PR:EMAIL WW571@mail.ustc.edu.cn PR:PHONE 18604523231 #STUDY ST:STUDY_TITLE Micropeptide hSPAR, a glutamine regulator, suppresses tumor growth via ST:STUDY_TITLE TRIM21-P27KIP1-mTOR pathway - human MDA-MB-231 breast cancer cell ST:STUDY_SUMMARY hSPAR, a microprotein, is capable of specifically inhibiting the mTOR signaling ST:STUDY_SUMMARY activity and cell proliferation in MDA-MB-231 cells. Glutamine, an essential ST:STUDY_SUMMARY amino acid, plays a crucial role in regulating the mTOR signal. Our data ST:STUDY_SUMMARY indicate that in MDA-MB-231 cells, overexpressing hSPAR inhibits glutamine ST:STUDY_SUMMARY uptake, consequently suppressing the activation of the mTOR signaling. ST:INSTITUTE University of Science and Technology of China ST:LAST_NAME Wang ST:FIRST_NAME Wei ST:ADDRESS Division of Life Sciences and Medicine, 443 Huangshan Road, Hefei city, Anhui ST:ADDRESS Province ST:EMAIL WW571@mail.ustc.edu.cn ST:PHONE 18604523231 #SUBJECT SU:SUBJECT_TYPE Human 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 - Ctrl4 Sample source:MDA-MB-231 breast cancer cell | Transfection:Ctrl RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206743a.mzML SUBJECT_SAMPLE_FACTORS - Ctrl5 Sample source:MDA-MB-231 breast cancer cell | Transfection:Ctrl RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206744a.mzML SUBJECT_SAMPLE_FACTORS - Ctrl6 Sample source:MDA-MB-231 breast cancer cell | Transfection:Ctrl RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206745a.mzML SUBJECT_SAMPLE_FACTORS - ATG4 Sample source:MDA-MB-231 breast cancer cell | Transfection:ATG RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206746a.mzML SUBJECT_SAMPLE_FACTORS - ATG5 Sample source:MDA-MB-231 breast cancer cell | Transfection:ATG RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206747a.mzML SUBJECT_SAMPLE_FACTORS - ATG6 Sample source:MDA-MB-231 breast cancer cell | Transfection:ATG RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206748a.mzML SUBJECT_SAMPLE_FACTORS - hsPAR4 Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206749a.mzML SUBJECT_SAMPLE_FACTORS - hsPAR5 Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206750a.mzML SUBJECT_SAMPLE_FACTORS - hsPAR6 Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206751a.mzML SUBJECT_SAMPLE_FACTORS - hsPAR_C4 Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR_C RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206752a.mzML SUBJECT_SAMPLE_FACTORS - hsPAR_C5 Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR_C RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206753a.mzML SUBJECT_SAMPLE_FACTORS - hsPAR_C6 Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR_C RAW_FILE_NAME(Raw_File_Name)=MWY-23-2561-a_12_WH6500-6_Amide-AA-02_V1.0_WQL_20231010-T23206754a.mzML #COLLECTION CO:COLLECTION_SUMMARY The MDA-MB-231 breast cancer cells were cultured in Dulbecco’s modified CO:COLLECTION_SUMMARY Eagle’s medium (DMEM), containing 10% fetal bovine serum and 1% CO:COLLECTION_SUMMARY penicillin-streptomycin under humidified atmosphere of 5% CO2 at 37°C. Ce11s CO:COLLECTION_SUMMARY were counted, washed with cold PBS and then flash-frozen in liquid N2. CO:SAMPLE_TYPE Breast cancer cells #TREATMENT TR:TREATMENT_SUMMARY MDA-MB-231 breast cancer cells were transfected with empty vector, ΔATG1+2, TR:TREATMENT_SUMMARY hSPAR or hSPAR-C for 48 hours. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The sample was thawed on ice, 100 μL of ultrapure water extract (containing SP:SAMPLEPREP_SUMMARY protease inhibitors, PMSF and EDTA) was added to resuspend the cell pellet. SP:SAMPLEPREP_SUMMARY Divide 50 μL cell suspension and add 200 µL of methanol (precooled at -20°C) SP:SAMPLEPREP_SUMMARY and vortexed for 2 min under the condition of 2500 rpm. The sample was frozen in SP:SAMPLEPREP_SUMMARY liquid nitrogen for 5 min, removed on ice for 5 min, after that, the sample was SP:SAMPLEPREP_SUMMARY vortexed for 2 min.The previous step was repeated for 3 times. The sample was SP:SAMPLEPREP_SUMMARY centrifuged at 12000 rpm for 10 min at 4°C. Take 200 μL of supernatant into a SP:SAMPLEPREP_SUMMARY new centrifuge tube and place the supernatant in -20°C refrigerator for 30 min. SP:SAMPLEPREP_SUMMARY Then the supernatant was centrifuged at 12000 rpm for 10 min at 4°C. After SP:SAMPLEPREP_SUMMARY centrifugation, transfer 180 μL of supernatant through Protein Precipitation SP:SAMPLEPREP_SUMMARY Plate for further LC-MS analysis. The remaining 50 μL cell suspension was SP:SAMPLEPREP_SUMMARY frozen and thawed for 3 times, centrifuged at 12,000 rpm for 10 min, and the SP:SAMPLEPREP_SUMMARY supernatant was taken to determine the protein concentration by BCA Protein SP:SAMPLEPREP_SUMMARY Assay kit. SP:SAMPLEPREP_PROTOCOL_FILENAME USTC_Protocol.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME SCIEX ExionLC AD CH:COLUMN_NAME Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um) CH:SOLVENT_A 100% water; 2 mM ammonium acetate; 0.04% formic acid CH:SOLVENT_B 100% acetonitrile; 2 mM ammonium acetate; 0.04% formic acid CH:FLOW_GRADIENT The gradient was started at 90% B at 0-1.2 min, decreased to 60% B at 9 min, 40% CH:FLOW_GRADIENT B at 10-11 min, finaly ramped back to 90% B at 11.01-15 min CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 40°C CH:METHODS_FILENAME USTC_Protocol.pdf #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE USTC_Protocol.pdf #MS MS:INSTRUMENT_NAME AB SCIEX QTRAP 5500 MS:INSTRUMENT_TYPE QTRAP MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS - #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS ng/mg of cells MS_METABOLITE_DATA_START Samples Ctrl4 Ctrl5 Ctrl6 ATG4 ATG5 ATG6 hsPAR4 hsPAR5 hsPAR6 hsPAR_C4 hsPAR_C5 hsPAR_C6 Factors Sample source:MDA-MB-231 breast cancer cell | Transfection:Ctrl Sample source:MDA-MB-231 breast cancer cell | Transfection:Ctrl Sample source:MDA-MB-231 breast cancer cell | Transfection:Ctrl Sample source:MDA-MB-231 breast cancer cell | Transfection:ATG Sample source:MDA-MB-231 breast cancer cell | Transfection:ATG Sample source:MDA-MB-231 breast cancer cell | Transfection:ATG Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR_C Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR_C Sample source:MDA-MB-231 breast cancer cell | Transfection:hsPAR_C Phosphorylethanolamine 8127.897866 6161.500481 7498.836722 8179.963444 7165.229925 7073.99741 67370.46407 63427.68257 52265.77976 53480.88486 59064.80695 62402.73733 Anserine 119.5364727 114.1004956 NA 108.5639554 NA 107.7391283 NA NA NA NA NA NA Ethanolamine 137.8299478 123.0642001 103.5298497 142.6997855 146.6473791 116.6120901 293.6711256 349.0149276 376.3288009 425.4399999 437.6160703 466.683817 Cys 327.420637 370.9861909 378.3696505 349.218778 366.0851305 341.5685744 NA 355.059711 NA 314.140629 278.1909001 386.6723013 γ-Glutamate-Cysteine 308.9223284 235.3955631 209.8178159 234.9890359 267.8855098 212.757212 135.3999714 266.5795556 128.9181577 162.1186394 NA NA L-Pipecolic-Acid 3.073135797 2.739290547 2.839725463 2.052810716 2.654286888 2.296054053 NA NA NA 2.463630404 NA NA Urea 690.8919745 861.0819557 982.318813 1086.954444 1005.644222 890.2561903 NA 1645.870024 NA NA NA NA argininosuccinic-acid 2779.706018 1922.357101 1822.654959 1407.980054 1220.969792 1683.658757 3859.039245 NA NA NA 3642.74777 NA 5-Hydroxy-Tryptamine 2356.218458 2031.522696 2872.477196 2777.475519 1947.161755 2381.405853 NA NA NA NA NA NA 5-Hydroxylysine 84.72133264 NA 87.90264585 75.76728209 NA NA NA NA NA NA NA NA Beta-Alanine 1837.529338 1645.992575 1996.713892 1633.211108 1859.288105 1676.098458 NA NA NA NA NA NA N-Acetylneuraminic-Acid 4201.561389 5226.203834 5383.327187 4929.501642 5231.623727 5264.632097 NA NA NA NA NA NA TRP-GLU 1162.717946 618.1388439 846.2410726 NA 588.8128254 528.3389156 NA NA NA NA NA NA Glycyl-L-Proline 11.44188567 10.44061498 11.62359595 10.96682493 10.46221842 10.60562985 25.39889583 30.21638274 28.77122669 NA 22.4765088 35.91129808 N,N-Dimethylglycine NA 1.580461067 0.907322754 2.241150016 2.60966239 1.403905577 NA NA NA NA NA NA 3-N-Methyl-L-Histidine NA 35.24738555 NA NA 32.81127503 34.29015746 NA NA NA NA NA NA Homo-Arg 134.0877083 133.5880528 137.8769824 128.0072247 123.3021374 133.4956532 345.9178342 NA 354.5803715 391.4344025 315.2503822 NA L-Homocitrulline 15.7260364 NA 14.41532961 NA NA 9.672330797 NA NA NA NA NA NA 1-Methylhistidine 179.5078165 169.7572659 174.3906572 157.2587191 164.4823363 153.3104902 47.00990692 47.11272725 42.84101239 37.53473404 33.68507191 48.62470582 L-Homocystine NA NA NA NA NA NA NA NA NA NA NA NA Nα-Acetyl-L-glutamine NA NA NA NA NA NA NA NA NA NA NA NA Nα-Acetyl-L-Arginine NA NA NA NA NA NA NA NA NA NA NA NA Nicotinuric-Acid NA NA NA NA NA NA NA NA NA NA NA NA N8-Acetylspermidine NA NA NA NA NA NA NA NA NA NA NA NA N-Acetyl-L-Tyrosine NA NA NA NA NA NA NA NA NA NA NA NA L-Theanine NA NA NA NA NA NA NA NA NA NA NA NA Homoserine NA NA NA NA NA NA NA NA NA NA NA NA D-Homocysteine NA NA NA NA NA NA NA NA NA NA NA NA 3-Chloro-L-Tyrosine NA NA NA NA NA NA NA NA NA NA NA NA 3-Aminoisobutanoic-Acid NA NA NA NA NA NA NA NA NA NA NA NA (5-L-Glutamyl)-L-Amino-Acid NA NA NA NA NA NA NA NA NA NA NA NA S-Sulfo-L-Cysteine NA NA NA NA NA NA NA NA NA NA NA NA S-(5-Adenosyl)-L-Homocysteine 631.4706893 610.2422225 622.0461475 525.8170719 646.1607469 567.116398 445.863494 554.2519455 410.5793013 388.3582752 458.7758837 487.1544873 3-Iodo-L-Tyrosine NA NA NA NA NA NA NA NA 37.59827344 NA NA NA O-Phospho-L-Serine 922.9004649 853.664161 960.5752682 873.4547746 795.6498457 913.7417516 1224.54222 NA 1670.180595 NA 976.9637068 NA L-tyrosine-methyl-ester 9.130707571 7.368327457 8.775168741 6.894321622 8.151866684 6.342626878 6.412644494 6.730223613 7.304186316 7.620049619 5.418762415 8.161138311 Methionine-Sulfoxide 29.89246112 29.18454404 40.37633609 23.60238438 24.84090439 26.62308552 144.1208001 148.9096224 179.9706889 277.9925818 137.8859193 217.7108602 Met 365.5806625 354.7060835 389.7384079 358.7993025 345.2194674 347.8201903 1420.171444 1648.767412 1632.116644 1555.14123 1726.60259 1643.479783 Lys 3325.518319 3151.575253 2886.839883 2820.610328 2994.076217 3082.126224 3217.716098 3183.625873 3467.249988 2854.141595 4563.148256 3814.673809 N-Acetylaspartate 12742.14127 12300.21938 11775.32361 11965.2862 10883.43014 10179.21349 6346.544506 10057.45948 5767.651214 7740.852093 9477.628707 5975.723243 L-Ornithine 3644.678872 3225.385755 3390.675712 3116.022306 3502.132554 3549.018917 10171.27828 10008.88231 9507.688932 8469.479391 11866.11789 11695.92375 L-Cystathionine 10483.85165 9339.003568 9787.611936 9111.194277 8273.955748 8978.098577 1106.175305 850.2053174 733.7516212 488.9435282 627.3671737 646.3501297 L-Citrulline 13524.88152 14909.42293 14191.89669 14943.45885 13487.87809 12134.0593 38056.89674 45923.41135 45866.23806 36742.90223 46447.62581 44633.78601 Ile 3202.641219 2996.123078 3086.391082 2667.879963 2903.966394 2807.523774 1012.35702 1278.733866 1258.341343 1205.195301 1313.984762 1465.511992 N-Glycyl-L-Leucine 56.22910429 45.96265414 58.38161617 40.33847674 52.11844488 49.90394399 146.7383857 142.5282473 122.7192002 104.6985922 116.8967354 130.8805963 His 1033.411604 904.1005721 995.2800859 885.3572027 995.3835561 953.1558908 1159.789015 999.4976349 884.6332399 843.4830909 1078.172518 1198.741791 Glu 323014.4147 297494.9063 330228.81 312534.4129 306434.7432 271508.6957 139178.7907 177041.2727 151503.8663 121529.4567 125606.7665 154806.0121 Gln 55764.66154 50225.93406 59233.88236 46250.45786 52819.07917 47958.38276 4182.919402 5507.548395 5278.655939 3770.405186 4298.892877 4208.507355 D-Alanyl-D-Alanine 30.43688101 21.00682724 27.13874677 23.75835386 26.61143823 25.8440652 113.8377446 86.66638303 97.49666668 79.36146934 76.64463244 127.1047604 Asp-Phe 17.00923788 19.82939586 17.9483101 21.52368208 16.05591404 20.14399745 35.39300528 35.1247753 28.59731989 24.47046228 45.12018158 58.64318867 Asp 3147.232599 2001.265902 1486.568096 615.3001544 2568.896774 2072.61626 7993.59837 9879.605585 7687.950506 8197.150547 7366.522499 9165.499477 Asn 1832.36247 1642.099956 1688.691358 1523.519341 1606.541282 1352.917769 1840.641716 2570.699011 2123.490705 2091.437683 2156.27749 2357.054124 Arg 67.96042664 64.18241151 65.6125978 62.57529963 58.37339442 69.52181535 472.3360989 383.9732437 393.9916301 351.3509306 522.1114193 409.0206518 Ala 14367.69092 12409.64907 12783.15151 12002.38106 12630.01913 11649.16116 4202.191771 4933.730647 5151.988894 4408.430629 4610.489851 4865.355741 Gly 9654.517529 8735.325979 9313.030315 9133.429817 9508.995614 9172.931027 3841.266925 5315.488682 3939.647511 4274.135057 4879.091728 4175.141884 N6-Acetyl-L-Lysine 10.3076718 10.58967132 9.268728338 9.333303804 9.642028068 9.182215605 27.01170945 12.60974714 33.86814734 15.17248424 42.20156639 16.5034367 Leu 5015.820683 4816.617552 4841.379158 4302.181468 4690.232593 4646.975541 3468.635173 4487.802778 4252.216086 3791.822209 4615.218136 4939.997603 Pro 13438.80541 12243.83954 12599.62162 12143.27742 12340.99882 11444.78991 3670.456991 5028.373655 4471.413912 3797.391382 4068.998333 4417.700015 L-Carnosine 172.1671813 168.9144391 175.6177955 151.4935139 142.7261621 157.8975139 415.8886007 470.6999734 424.8049849 478.9040651 361.7197405 591.7787383 Glutathione-Oxidized 10845.02609 10451.40868 13569.91424 10419.50994 9982.628752 9380.430294 9893.182231 7609.635201 4528.834067 3508.2541 4589.579667 4729.799791 glycylphenylalanine 7.090749776 9.05016888 11.12600999 9.909973983 9.322017979 6.18061099 111.7363715 128.0540516 123.1690866 79.7722426 84.38521815 96.66539957 Val 3610.19089 3421.463562 3461.743967 3140.780725 3313.195795 3249.624434 1318.412681 1753.017154 1776.527156 1568.182066 1799.893958 2015.902655 Tyr 1266.665206 1180.71542 1231.10458 1073.121372 1198.301338 1122.947612 1740.203649 2371.16906 1980.914682 2034.665753 2186.109743 2502.182288 Trp 1317.977155 1296.437611 1312.953865 1192.498428 1269.571905 1228.449157 1577.229841 2304.5029 1964.33037 1798.394503 2214.226891 2978.940895 Trimethylamine-N-Oxide 1.781660418 1.985718388 1.665266357 1.548769507 1.446625289 1.392344067 0.805399165 1.122616007 2.027593336 2.268156154 0.920268329 2.593883818 Phe 2240.452993 2115.690553 2178.476994 1965.065854 2067.450007 2042.021365 3421.644506 4249.827151 3996.597284 3751.866193 4302.82822 4207.804679 Trans-4-Hydroxy-L-Proline 901.7105517 792.9427364 828.9914261 765.2478707 762.9275228 701.7416749 359.0921781 426.4745138 329.1898446 327.4408618 268.153131 390.5453949 Sarcosine NA NA NA NA NA NA NA NA NA NA NA NA Ser 2533.549055 2211.662578 2521.895438 2484.479518 2366.810135 2252.770234 2216.58988 3054.780194 2504.593879 2016.545382 2253.988545 2603.781775 Thr 8989.982336 8702.807102 8827.559494 8482.110226 8414.161763 8022.803918 3950.509628 4310.586088 4167.513799 4046.845501 3892.700818 4193.374229 α-Aminoadipic-acid 1541.303236 1377.183636 1507.997945 1290.131691 1435.009067 1232.328785 489.322524 517.1443898 630.1030352 NA 566.5573063 506.7456494 Kinurenine 7.290600553 6.120116995 6.640993723 6.213753743 6.329623839 6.385391202 NA NA NA NA NA NA Creatine-Phosphate NA NA NA NA NA NA NA NA NA NA 22207.99502 38288.71585 Creatine 33096.03445 31341.33101 32344.40933 29111.13253 28623.3995 28688.57255 14594.27075 17560.45731 14741.05384 13136.33462 11926.08783 14684.29726 N'-Formylkynurenine NA NA NA NA NA NA NA NA NA NA NA NA Kynurenic-Acid NA NA NA NA NA NA NA NA NA NA NA NA 5-Aminovaleric-Acid 64.01507977 59.28127014 63.146182 50.18358281 45.37124062 49.82414083 NA NA NA NA NA NA (S)-β-Aminoisobutyric-Acid NA NA NA NA NA NA NA NA NA NA NA NA 6-Aminocaproic-Acid NA NA NA NA NA NA NA NA NA NA NA NA 3-Hydroxyhippuric-Acid NA NA NA NA NA NA NA NA NA NA NA NA γ-Aminobutyric-Acid 2352.950327 2410.163225 2575.283767 1812.48515 2160.884405 1973.61392 NA NA 1368.481195 NA 582.66872 NA 1,3,7-Trimethyluric-Acid NA NA NA NA NA NA NA NA NA NA NA NA 2-Aminobutyric-acid 86.75321217 81.91897878 62.95580128 68.65661294 73.64763766 56.68080174 NA NA NA NA NA NA 4-Acetamidobutyric-Acid 27.27266334 21.78454742 22.66215581 18.00993224 17.93924594 20.43524582 NA NA NA NA NA NA Guanidinoethyl-Sulfonate NA NA NA NA NA NA NA NA NA NA NA NA P-Aminohippuric-Acid NA NA NA NA NA NA NA NA NA NA NA NA MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Compounds Class Q1 (Da) Q3 (Da) Molecular Weight Ion mode Ionization model Formula cpd_ID HMDB CAS kegg_map Phosphorylethanolamine Phosphorylethanolamine - 142.026 44.049 141.06 Positive [M+H]+ C2H8NO4P - - 1071-23-4 - Anserine Anserine - 241 170 240.26 Positive [M+H]+ C10H16N4O3 - - 584-85-0 - Ethanolamine Ethanolamine - 62 44 61.08 Positive [M+H]+ C2H7NO - - 141-43-5 - Cys L-Cysteine Amino Acid metabolomics 122.02 59 121.02 Positive [M+H]+ C3H7NO2S C00097 HMDB00574 52-90-4 ko00260,ko00270,ko00430,ko00470,ko00480,ko00730,ko00770,ko00920,ko00970,ko01100,ko01200,ko01230,ko01240,ko04122,ko04216,ko04974,ko05230 γ-Glutamate-Cysteine γ-Glutamate-Cysteine Amino Acid metabolomics 251.07 130.05 250.06 Positive [M+H]+ C8H14N2O5S C00669 HMDB01049 636-58-8 ko00480,ko01100,ko01240,ko04216 L-Pipecolic-Acid L-Pipecolic Acid Amino Acid metabolomics 130 84 129.08 Positive [M+H]+ C6H11NO2 C00408 HMDB00716 3105-95-1 ko00310,ko01100 Urea Urea Amino Acid metabolomics 61 44 60.03 Positive [M+H]+ CH4N2O C00086 HMDB00294 57-13-6 ko00220,ko00230,ko00240,ko00330,ko00780,ko01100,ko02010,ko05120 argininosuccinic-acid argininosuccinic acid Amino Acid metabolomics 291.5 176.1 290.27 Positive [M+H]+ C10H18N4O6 - - 2387-71-5 - 5-Hydroxy-Tryptamine 5-Hydroxy-Tryptamine Amino Acid metabolomics 177.1 160 176.21 Positive [M+H]+ C10H12N20 - - 153-98-0(50-67-9) - 5-Hydroxylysine 5-Hydroxylysine Amino Acid metabolomics 163.1 128.07 162.18 Positive [M+H]+ C6H14N2O3 - - 13204-98-3(1190-94-9) - Beta-Alanine Beta-Alanine Amino Acid metabolomics 90.2 30 89.05 Positive [M+H]+ C3H7NO2 C00099 HMDB00056 107-95-9 ko00240,ko00410,ko00640,ko00770,ko01100,ko01240,ko04080,ko04974 N-Acetylneuraminic-Acid N-Acetylneuraminic Acid Amino Acid metabolomics 310.11 274.09 309.11 Positive [M+H]+ C11H19NO9 C19910 HMDB00230 131-48-6 - TRP-GLU L-Tryptophyl-L-glutamic acid Amino Acid metabolomics 334 159 333.13 Positive [M+H]+ C16H19N3O5 - HMDB29082 36099-95-3 - Glycyl-L-Proline Glycyl-L-Proline Amino Acid metabolomics 173.1 116.071 172.09 Positive [M+H]+ C7H12N2O3 - HMDB00721 704-15-4 - N,N-Dimethylglycine N,N-Dimethylglycine Amino Acid metabolomics 104 58 103.06 Positive [M+H]+ C4H9NO2 C01026 HMDB00092 1118-68-9 ko00260,ko01100 3-N-Methyl-L-Histidine 3-N-Methyl-L-Histidine Amino Acid metabolomics 170 96 169.18 Positive [M+H]+ C7H11N3O2 C01152 HMDB00479 368-16-1 ko00340,ko01100 Homo-Arg Homo-L-arginine Amino Acid metabolomics 189 144 188.13 Positive [M+H]+ C7H16N4O2 C01924 HMDB00670 156-86-5 - L-Homocitrulline L-Homocitrulline Amino Acid metabolomics 190.1 127 189.11 Positive [M+H]+ C7H15N3O3 C02427 HMDB00679 1190-49-4 - 1-Methylhistidine 1-Methylhistidine Amino Acid metabolomics 170.1 124 169.09 Positive [M+H]+ C7H11N3O2 C01152 HMDB00001 332-80-9 ko00340,ko01100 L-Homocystine L-Homocystine Amino Acid metabolomics 269 136 268.06 Positive [M+H]+ C8H16N2O4S2 C01817 HMDB00676 626-72-2 ko00270 Nα-Acetyl-L-glutamine Nα-Acetyl-L-glutamine Amino Acid metabolomics 189.1 130.1 188.18 Positive [M+H]+ C7H12N2O4 - HMDB06029 2490-97-3 - Nα-Acetyl-L-Arginine Nα-Acetyl-L-Arginine Amino Acid metabolomics 217.1 158 216.12 Positive [M+H]+ C8H16N4O3 - HMDB04620 155-84-0 - Nicotinuric-Acid Nicotinuric Acid Amino Acid metabolomics 181.1 135 180.05 Positive [M+H]+ C8H8N2O3 C05380 HMDB03269 583-08-4 ko00760 N8-Acetylspermidine N8-Acetylspermidine Amino Acid metabolomics 190.18 173.15 189.18 Positive [M+H]+ C9H23N3O C01029 HMDB02189 34450-15-2(13431-24-8) - N-Acetyl-L-Tyrosine N-Acetyl-L-Tyrosine Amino Acid metabolomics 224 136 223.09 Positive [M+H]+ C11H13NO4 C01657 HMDB00866 537-55-3 - L-Theanine L-Theanine Amino Acid metabolomics 175 158 174.10 Positive [M+H]+ C7H14N2O3 - HMDB34365 3081-61-6 - Homoserine Homoserine Amino Acid metabolomics 120.065 74.06 119.12 Positive [M+H]+ C4H9NO3 - - 1927-25-9 - D-Homocysteine D-Homocysteine Amino Acid metabolomics 136 90.036 135.04 Positive [M+H]+ C4H9NO2S C00155 HMDB00742 454-29-5 ko00270,ko00920,ko01100,ko01230,ko01523,ko04621 3-Chloro-L-Tyrosine 3-Chloro-L-Tyrosine Amino Acid metabolomics 216.1 170.1 215.04 Positive [M+H]+ C9H10ClNO3 - HMDB01885 7423-93-0 - 3-Aminoisobutanoic-Acid 3-Aminoisobutanoic Acid Amino Acid metabolomics 104.07 86.05 103.06 Positive [M+H]+ C4H9NO2 C05145 HMDB03911 144-90-1 - (5-L-Glutamyl)-L-Amino-Acid (5-L-Glutamyl)-L-Amino Acid Amino Acid metabolomics 219.09 202 218.09 Positive [M+H]+ C8H14N2O5 C03740 HMDB06248 5875-41-2 ko00480,ko01100 S-Sulfo-L-Cysteine S-Sulfo-L-Cysteine Amino Acid metabolomics 201.98 120 200.98 Positive [M+H]+ C3H7NO5S2 C05824 HMDB00731 1637-71-4 ko00270,ko01100,ko01230 S-(5-Adenosyl)-L-Homocysteine S-(5-Adenosyl)-L-Homocysteine Amino Acid metabolomics 385 136.06 384.12 Positive [M+H]+ C14H20N6O5S C00021 HMDB00939 979-92-0 ko00270,ko01100,ko01230,ko01240,ko05208 3-Iodo-L-Tyrosine 3-Iodo-L-Tyrosine Amino Acid metabolomics 308.2 261.97 306.97 Positive [M+H]+ C9H10INO3 C02515 HMDB00021 70-78-0 ko00350,ko01100,ko04918 O-Phospho-L-Serine O-Phospho-L-Serine Amino Acid metabolomics 186 88.2 185.01 Positive [M+H]+ C3H8NO6P C01005 HMDB00272 407-41-0 ko00260,ko00270,ko00970,ko01100,ko01200,ko01230 L-tyrosine-methyl-ester L-tyrosine methyl ester Amino Acid metabolomics 196.2 136.3 195.09 Positive [M+H]+ C10H13NO3 C03404 HMDB29217 1080-06-4 - Methionine-Sulfoxide Methionine Sulfoxide Amino Acid metabolomics 166 74 165.05 Positive [M+H]+ C5H11NO3S - HMDB02005 62697-73-8 - Met L-Methionine Amino Acid metabolomics 150.05 104 149.05 Positive [M+H]+ C5H11NO2S C00073 HMDB00696 63-68-3 ko00270,ko00470,ko00970,ko01100,ko01210,ko01230,ko01240,ko01523,ko04974,ko04978,ko05230 Lys L-Lysine Amino Acid metabolomics 147.069 84 146.11 Positive [M+H]+ C6H14N2O2 C00047 HMDB00182 56-87-1 ko00310,ko00470,ko00780,ko00970,ko01100,ko01210,ko01230,ko02010,ko04974 N-Acetylaspartate N-Acetylaspartate Amino Acid metabolomics 176 134 175.05 Positive [M+H]+ C6H9NO5 C01042 HMDB00812 997-55-7 ko00250,ko01100 L-Ornithine L-Ornithine Amino Acid metabolomics 133.09 70 132.16 Positive [M+H]+ C5H13ClN2O2 C00077 HMDB00214 3184-13-2 ko00220,ko00330,ko00470,ko00480,ko01100,ko01210,ko01230,ko02010 L-Cystathionine L-Cystathionine Amino Acid metabolomics 223.07 134 222.07 Positive [M+H]+ C7H14N2O4S C02291 HMDB00099 56-88-2 ko00260,ko00270,ko01100,ko01230,ko01240 L-Citrulline L-Citrulline Amino Acid metabolomics 176.1 113 175.10 Positive [M+H]+ C6H13N3O3 C00327 HMDB00904 372-75-8 ko00220,ko01100,ko01230 Ile L-Isoleucine Amino Acid metabolomics 132.1 86.1 131.10 Positive [M+H]+ C6H13NO2 C00407 HMDB00172 73-32-5 ko00280,ko00290,ko00970,ko01100,ko01210,ko01230,ko02010,ko04974,ko04978,ko05131,ko05230 N-Glycyl-L-Leucine N-Glycyl-L-Leucine Amino Acid metabolomics 189.1 86 188.12 Positive [M+H]+ C8H16N2O3 C02155 HMDB00759 869-19-2 - His L-Histidine Amino Acid metabolomics 156.07 110.07 155.07 Positive [M+H]+ C6H9N3O2 C00135 HMDB00177 71-00-1 ko00340,ko00410,ko00470,ko00970,ko01100,ko01230,ko02010,ko04974,ko05230 Glu L-Glutamic acid Amino Acid metabolomics 148.06 84 147.05 Positive [M+H]+ C5H9NO4 C00025 HMDB00148 56-86-0 ko00220,ko00250,ko00330,ko00340,ko00430,ko00470,ko00480,ko00524,ko00630,ko00650,ko00860,ko00910,ko00970,ko01100,ko01200,ko01210,ko01230,ko01240,ko02010,ko04068,ko04072,ko04080,ko04216,ko04540,ko04713,ko04720,ko04721,ko04723,ko04724,ko04727,ko04730,ko04742,ko04964,ko04974,ko05014,ko05016,ko05017,ko05022,ko05030,ko05031,ko05033,ko05034,ko05230 Gln L-Glutamine Amino Acid metabolomics 147.069 84 146.07 Positive [M+H]+ C5H10N2O3 C00064 HMDB00641 56-85-9 ko00220,ko00230,ko00240,ko00250,ko00470,ko00630,ko00750,ko00910,ko00970,ko01100,ko01230,ko01232,ko01240,ko02010,ko04724,ko04727,ko04964,ko04974,ko04978,ko05230 D-Alanyl-D-Alanine D-Alanyl-D-Alanine Amino Acid metabolomics 161.09 44.04 160.09 Positive [M+H]+ C6H12N2O3 C00993 HMDB03459 923-16-0 ko00470,ko01100 Asp-Phe L-α-Aspartyl-L-phenylalanine Amino Acid metabolomics 281.3 166 280.11 Positive [M+H]+ C13H16N2O5 - HMDB00706 13433-09-5 - Asp L-Aspartate Amino Acid metabolomics 134.04 74 133.10 Positive [M+H]+ C4H7NO4 C00049 HMDB00191 56-84-8 ko00220,ko00250,ko00260,ko00270,ko00340,ko00410,ko00470,ko00760,ko00770,ko00970,ko01100,ko01200,ko01210,ko01230,ko01240,ko02010,ko04080,ko04974,ko05230 Asn L-Asparagine Anhydrous Amino Acid metabolomics 133.06 74 132.05 Positive [M+H]+ C4H8N2O3 C00152 HMDB00168 70-47-3 ko00250,ko00970,ko01100,ko01230,ko04974,ko04978,ko05230 Arg L-Arginine Amino Acid metabolomics 175.1 70.06 174.11 Positive [M+H]+ C6H14N4O2 C00062 HMDB00517 74-79-3 ko00220,ko00330,ko00470,ko00970,ko01100,ko01230,ko02010,ko04150,ko04974,ko05014,ko05022,ko05142,ko05146,ko05230 Ala L-Alanine Amino Acid metabolomics 90.05 44.05 89.05 Positive [M+H]+ C3H7NO2 C00041 HMDB00161 56-41-7 ko00250,ko00270,ko00430,ko00450,ko00470,ko00970,ko01100,ko01200,ko01230,ko02010,ko04122,ko04974,ko04978,ko05230 Gly Glycine Amino Acid metabolomics 76.03 30 75.03 Positive [M+H]+ C2H5NO2 C00037 HMDB00123 56-40-6 ko00120,ko00230,ko00260,ko00310,ko00440,ko00470,ko00480,ko00630,ko00730,ko00785,ko00860,ko00970,ko01100,ko01200,ko01230,ko01240,ko02010,ko04080,ko04721,ko04974,ko04978,ko05230 N6-Acetyl-L-Lysine N6-Acetyl-L-Lysine Amino Acid metabolomics 189.1 126 188.12 Positive [M+H]+ C8H16N2O3 C02727 HMDB00206 692-04-6 ko00310,ko01100 Leu L-Leucine Amino Acid metabolomics 132.1 86.1 131.10 Positive [M+H]+ C6H13NO2 C00123 HMDB00687 61-90-5 ko00280,ko00290,ko00970,ko01100,ko01210,ko01230,ko02010,ko04150,ko04974,ko04978,ko05131,ko05230 Pro L-Proline Amino Acid metabolomics 116.1 70.1 115.06 Positive [M+H]+ C5H9NO2 C00148 HMDB00162 147-85-3 ko00330,ko00470,ko00970,ko01100,ko01230,ko02010,ko04974,ko04978,ko05230 L-Carnosine L-Carnosine Amino Acid metabolomics 227.2 156 226.11 Positive [M+H]+ C9H14N4O3 C00386 HMDB00033 305-84-0 ko00340,ko00410,ko01100 Glutathione-Oxidized Glutathione Oxidized Amino Acid metabolomics 613.152 484 612.15 Positive [M+H]+ C20H32N6O12S2 - - 121-24-4 - glycylphenylalanine glycylphenylalanine Amino Acid metabolomics 223.2 120.2 222.10 Positive [M+H]+ C11H14N2O3 - HMDB28848 3321-03-7 - Val L-Valine Amino Acid metabolomics 118.1 72.1 117.08 Positive [M+H]+ C5H11NO2 C00183 HMDB00883 72-18-4 ko00280,ko00290,ko00770,ko00970,ko01100,ko01210,ko01230,ko01240,ko02010,ko04974,ko04978,ko05230 Tyr L-Tyrosine Amino Acid metabolomics 182.08 136.07 181.19 Positive [M+H]+ C9H11NO3 C00082 HMDB00158 60-18-4 ko00130,ko00350,ko00360,ko00400,ko00730,ko00970,ko01100,ko01210,ko01230,ko01240,ko04728,ko04916,ko04917,ko04974,ko05012,ko05030,ko05031,ko05034,ko05230 Trp L-Tryptophan Amino Acid metabolomics 205.1 118.06 204.09 Positive [M+H]+ C11H12N2O2 C00078 HMDB00929 73-22-3 ko00260,ko00380,ko00400,ko00970,ko01100,ko01210,ko01230,ko01240,ko04726,ko04974,ko04978,ko05143,ko05230 Trimethylamine-N-Oxide Trimethylamine N-Oxide Amino Acid metabolomics 76 58.1 75.07 Positive [M+H]+ C3H9NO C01104 HMDB00925 1184-78-7 ko01100 Phe L-Phenylalanine Amino Acid metabolomics 166.1 120.08 165.08 Positive [M+H]+ C9H11NO2 C00079 HMDB00159 63-91-2 ko00360,ko00400,ko00470,ko00970,ko01100,ko01210,ko01230,ko02010,ko04974,ko04978,ko05230 Trans-4-Hydroxy-L-Proline Trans-4-Hydroxy-L-Proline Amino Acid metabolomics 132.058 86 131.06 Positive [M+H]+ C5H9NO3 C01157 HMDB00725 51-35-4 ko00330,ko00470,ko01100,ko02010 Sarcosine Sarcosine Amino Acid metabolomics 90.2 44.1 89.05 Positive [M+H]+ C3H7NO2 C00213 HMDB00271 107-97-1 ko00260,ko00330,ko01100 Ser L-Serine Amino Acid metabolomics 106.04 60 105.04 Positive [M+H]+ C3H7NO3 C00065 HMDB00187 56-45-1 ko00260,ko00270,ko00470,ko00564,ko00600,ko00630,ko00920,ko00970,ko01100,ko01200,ko01230,ko01240,ko02010,ko04071,ko04974,ko04978,ko05230 Thr L-Threonine Amino Acid metabolomics 120.06 74 119.06 Positive [M+H]+ C4H9NO3 C00188 HMDB00167 72-19-5 ko00260,ko00290,ko00470,ko00860,ko00970,ko01100,ko01230,ko02010,ko04974,ko04978 α-Aminoadipic-acid α-Aminoadipic acid Organic Acid And Its Derivatives 162.07 98.06 161.16 Positive [M+H]+ C6H11NO4 - - 1118-90-7(542-32-5) - Kinurenine Kinurenine Organic Acid And Its Derivatives 209.1 146 208.09 Positive [M+H]+ C10H12N2O3 C00328 HMDB00684 343-65-7 ko00380,ko01100,ko01240,ko05143 Creatine-Phosphate Creatine Phosphate Organic Acid And Its Derivatives 212.04 114.06 211.11 Positive [M+H]+ C4H8N3Na2O5P - - 922-32-7 - Creatine Creatine Organic Acid And Its Derivatives 132.1 90 131.07 Positive [M+H]+ C4H9N3O2 C00300 HMDB00064 57-00-1 ko00260,ko00330,ko01100 N'-Formylkynurenine N'-Formylkynurenine Organic Acid And Its Derivatives 237.1 146.06 236.08 Positive [M+H]+ C11H12N2O4 - HMDB01200 1022-31-7 - Kynurenic-Acid Kynurenic Acid Organic Acid And Its Derivatives 190.043 144 189.04 Positive [M+H]+ C10H7NO3 C01717 HMDB00715 492-27-3 ko00380,ko01100 5-Aminovaleric-Acid 5-Aminovaleric Acid Organic Acid And Its Derivatives 118.2 55 117.08 Positive [M+H]+ C5H11NO2 C00431 HMDB03355 660-88-8 ko00310,ko00330,ko00470,ko01100 (S)-β-Aminoisobutyric-Acid (S)-β-Aminoisobutyric Acid Organic Acid And Its Derivatives 104.07 57.03 103.06 Positive [M+H]+ C4H9NO2 C03284 HMDB02166 4249-19-8 ko00280,ko01100 6-Aminocaproic-Acid 6-Aminocaproic Acid Organic Acid And Its Derivatives 132.095 69 131.10 Positive [M+H]+ C6H13NO2 C02378 HMDB01901 60-32-2 ko01100 3-Hydroxyhippuric-Acid 3-Hydroxyhippuric Acid Organic Acid And Its Derivatives 196.05 121 195.05 Positive [M+H]+ C9H9NO4 - HMDB06116 1637-75-8 - γ-Aminobutyric-Acid γ-Aminobutyric Acid Organic Acid And Its Derivatives 104.06 68.8 103.06 Positive [M+H]+ C4H9NO2 C00334 HMDB00112 56-12-2 ko00250,ko00330,ko00410,ko00650,ko00760,ko01100,ko04024,ko04080,ko04721,ko04723,ko04727,ko04742,ko04915,ko04929,ko05032,ko05033 1,3,7-Trimethyluric-Acid 1,3,7-Trimethyluric Acid Organic Acid And Its Derivatives 211.08 154.06 210.08 Positive [M+H]+ C8H10N4O3 - HMDB02123 5415-44-1 - 2-Aminobutyric-acid 2-Aminobutyric acid Organic Acid And Its Derivatives 104.1 58.14 103.12 Positive [M+H]+ C4H9NO2 - - 1492-24-6 - 4-Acetamidobutyric-Acid 4-Acetamidobutyric Acid Organic Acid And Its Derivatives 146.074 86 145.07 Positive [M+H]+ C6H11NO3 C02946 HMDB03681 3025-96-5 ko00330,ko01100 Guanidinoethyl-Sulfonate Guanidinoethyl Sulfonate Organic Acid And Its Derivatives 168.04 151.01 167.04 Positive [M+H]+ C3H9N3O3S C01959 HMDB03584 543-18-0 ko00430 P-Aminohippuric-Acid P-Aminohippuric Acid Organic Acid And Its Derivatives 195.069 120 194.07 Positive [M+H]+ C9H10N2O3 - HMDB01867 61-78-9 - METABOLITES_END #END