#METABOLOMICS WORKBENCH juliehaines_20241126_063750 DATATRACK_ID:5402 STUDY_ID:ST003607 ANALYSIS_ID:AN005927 PROJECT_ID:PR002231
VERSION             	1
CREATED_ON             	December 5, 2024, 7:47 pm
#PROJECT
PR:PROJECT_TITLE                 	Macrophages recycle phagocytosed bacteria to fuel immunometabolic responses
PR:PROJECT_SUMMARY               	Macrophages specialize in phagocytosis, a cellular process that eliminates
PR:PROJECT_SUMMARY               	extracellular matter, including microbes, through internalization and
PR:PROJECT_SUMMARY               	degradation. Despite the critical role of phagocytosis during bacterial
PR:PROJECT_SUMMARY               	infection, the fate of phagocytosed microbial cargo and its impact on host cell
PR:PROJECT_SUMMARY               	is poorly understood. Here, we reveal that ingested bacteria constitute an
PR:PROJECT_SUMMARY               	alternative nutrient source that skews immunometabolic host responses. Tracing
PR:PROJECT_SUMMARY               	stable isotope-labelled bacteria, we found that phagolysosomal degradation of
PR:PROJECT_SUMMARY               	bacteria provides carbon atoms and amino acids that are recycled into various
PR:PROJECT_SUMMARY               	metabolic pathways, including glutathione and itaconate biosynthesis, and
PR:PROJECT_SUMMARY               	satisfy macrophage bioenergetic needs. Metabolic recycling of
PR:PROJECT_SUMMARY               	microbially-derived nutrients is regulated by the nutrient sensing mTORC1 and
PR:PROJECT_SUMMARY               	intricately tied to microbial viability. Dead bacteria, as opposed to live ones,
PR:PROJECT_SUMMARY               	are enriched in cyclic- adenosine monophosphate (AMP), sustain the cellular AMP
PR:PROJECT_SUMMARY               	pool and subsequently activate AMP protein kinase (AMPK) to inhibit mTORC1.
PR:PROJECT_SUMMARY               	Consequently, killed bacteria strongly fuel metabolic recycling and support
PR:PROJECT_SUMMARY               	macrophage survival, but elicit decreased reactive oxygen species (ROS)
PR:PROJECT_SUMMARY               	production and a reduced IL-1β secretion compared to viable bacteria. These
PR:PROJECT_SUMMARY               	results reveal a novel insight into the fate of engulfed microbes and highlights
PR:PROJECT_SUMMARY               	a microbial viability-associated metabolite that triggers host metabolic and
PR:PROJECT_SUMMARY               	immune responses. Our findings hold promise for shaping immunometabolic
PR:PROJECT_SUMMARY               	intervention in various immune-related pathologies.
PR:INSTITUTE                     	University of Colorado Anschutz Medical Campus
PR:LABORATORY                    	Lab of Angelo D'Alessandro in collaboration with lab of Johan Garaude (INSERM,
PR:LABORATORY                    	Fr)
PR:LAST_NAME                     	Haines
PR:FIRST_NAME                    	Julie
PR:ADDRESS                       	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
PR:EMAIL                         	julie.haines@cuanschutz.edu
PR:PHONE                         	3037243339
#STUDY
ST:STUDY_TITLE                   	Global metabolomics and tracing of 13C-labeled metabolites of bone marrow
ST:STUDY_TITLE                   	derived macrophages following engulfment of dead 13C-labeled Citrobacter
ST:STUDY_TITLE                   	rodentium, Listeria innocua, Staphilococcus aureus, or Pseudomonas aeruginosa.
ST:STUDY_SUMMARY                 	Global metabolomics and 13C-tracing analysis of bone marrow derived macrophages
ST:STUDY_SUMMARY                 	exposed to heat-killed uniformly 13C-labelled Citrobacter rodentium, Listeria
ST:STUDY_SUMMARY                 	innocua, Staphilococcus aureus or Pseudomonas aeruginosa for 4h.
ST:INSTITUTE                     	University of Colorado Anschutz Medical Campus
ST:LAST_NAME                     	Haines
ST:FIRST_NAME                    	Julie
ST:ADDRESS                       	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
ST:EMAIL                         	julie.haines@cuanschutz.edu
ST:PHONE                         	3037243339
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Mus musculus
SU:GENDER                        	Not applicable
#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           	-	AA89-33_r67+	treatment:CTRL | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-33_r67
SUBJECT_SAMPLE_FACTORS           	-	AA89-34_r102+	treatment:CTRL | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-34_r102
SUBJECT_SAMPLE_FACTORS           	-	AA89-35_r39+	treatment:CTRL | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-35_r39
SUBJECT_SAMPLE_FACTORS           	-	AA89-36_r15+	treatment:CTRL | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-36_r15
SUBJECT_SAMPLE_FACTORS           	-	AA89-37_r55+	treatment:Killed-Citro | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-37_r55
SUBJECT_SAMPLE_FACTORS           	-	AA89-38_r8+	treatment:Killed-Citro | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-38_r8
SUBJECT_SAMPLE_FACTORS           	-	AA89-39_r42+	treatment:Killed-Citro | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-39_r42
SUBJECT_SAMPLE_FACTORS           	-	AA89-40_r32+	treatment:Killed-Citro | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-40_r32
SUBJECT_SAMPLE_FACTORS           	-	AA89-41_r53+	treatment:Killed-Listeria | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-41_r53
SUBJECT_SAMPLE_FACTORS           	-	AA89-42_r38+	treatment:Killed-Listeria | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-42_r38
SUBJECT_SAMPLE_FACTORS           	-	AA89-43_r65+	treatment:Killed-Listeria | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-43_r65
SUBJECT_SAMPLE_FACTORS           	-	AA89-44_r43+	treatment:Killed-Listeria | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-44_r43
SUBJECT_SAMPLE_FACTORS           	-	AA89-45_r105+	treatment:Killed_Staph | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-45_r105
SUBJECT_SAMPLE_FACTORS           	-	AA89-46_r70+	treatment:Killed_Staph | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-46_r70
SUBJECT_SAMPLE_FACTORS           	-	AA89-47_r81+	treatment:Killed_Staph | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-47_r81
SUBJECT_SAMPLE_FACTORS           	-	AA89-48_r80+	treatment:Killed_Staph | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-48_r80
SUBJECT_SAMPLE_FACTORS           	-	AA89-49_r108+	treatment:Killed-Pseudo | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-49_r108
SUBJECT_SAMPLE_FACTORS           	-	AA89-50_r36+	treatment:Killed-Pseudo | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-50_r36
SUBJECT_SAMPLE_FACTORS           	-	AA89-51_r33+	treatment:Killed-Pseudo | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-51_r33
SUBJECT_SAMPLE_FACTORS           	-	AA89-52_r12+	treatment:Killed-Pseudo | Sample source:murine bone marrow derived macrophages	RAW_FILE_NAME(raw file name)=AA89-52_r12
#COLLECTION
CO:COLLECTION_SUMMARY            	BMDMs were seeded at 2.5E6 cell/well in a non-tissue treated 6-well plate (2
CO:COLLECTION_SUMMARY            	wells per conditions) the day before the experiment. For metabolite extraction,
CO:COLLECTION_SUMMARY            	BMDMs were washed with cold PBS and harvested with 5 mM EDTA in PBS, frozen as
CO:COLLECTION_SUMMARY            	dry cell pellet, and stored at -80˚C until processing.
CO:SAMPLE_TYPE                   	Macrophages
#TREATMENT
TR:TREATMENT_SUMMARY             	Preparation of macrophages: Murine bone marrow-derived macrophages (BMDMs) were
TR:TREATMENT_SUMMARY             	generated in RPMI 1640 supplemented with M-CSF (30% mycoplasma-free L929 cell
TR:TREATMENT_SUMMARY             	supernatant, NCBI Biosample accession # SAMN00155972) and 10% FBS, plus 100
TR:TREATMENT_SUMMARY             	µg/ml penicillin, 100 µg/ml streptomycin, 10 mM HEPES, 1 nM sodium pyruvate
TR:TREATMENT_SUMMARY             	and 50 µM 2-mercaptoethanol (all from Gibco). BMDMs were used on day 5 to 7
TR:TREATMENT_SUMMARY             	after seeding. Period of differentiation of the cells, concentration of cells
TR:TREATMENT_SUMMARY             	when replating and time-lapse between replating and stimulation with bacteria
TR:TREATMENT_SUMMARY             	were important parameters to maintain metabolic backgrounds homogenous between
TR:TREATMENT_SUMMARY             	experiments. Preparation of viable and killed U-[13C]Bacteria: Citrobacter
TR:TREATMENT_SUMMARY             	rodentium, Staphylococus aureus, Listeria innocua, Pseudomonas aeruginosa were
TR:TREATMENT_SUMMARY             	grown overnight with shaking in LB, diluted 1/40, and grown until log-phase
TR:TREATMENT_SUMMARY             	[optical density at 600 nm (OD600) of 0.8-1.2]. Bacteria were washed with
TR:TREATMENT_SUMMARY             	phosphate buffer saline (PBS) to remove LB salts before addition to cells. For
TR:TREATMENT_SUMMARY             	labeling, 10 µl of an overnight cultured of bacteria was added to 20 ml of a
TR:TREATMENT_SUMMARY             	filtered M9 minimal medium salts (Life Technologies) supplemented with 1 mM
TR:TREATMENT_SUMMARY             	thiamine, 1 mM MgSO4, 0.1 M CaCl2, and 0.5% U-13C glucose (Campro Scientific).
TR:TREATMENT_SUMMARY             	Bacteria were grown for 72h, washed with PBS and subjected to heat-killing by
TR:TREATMENT_SUMMARY             	re-suspension in PBS and subsequently incubation at 60˚C for 60-90 min. For
TR:TREATMENT_SUMMARY             	antibiotic killing, bacteria were incubated for 6h to 12h with streptomycin or
TR:TREATMENT_SUMMARY             	gentamycin (50 µg/ml). Bacteria were kept at 4˚C until use. Efficient killing
TR:TREATMENT_SUMMARY             	was confirmed by overnight plating on LB-agar plates. Treatment of macrophages:
TR:TREATMENT_SUMMARY             	2E6 BMDMs were plated 12-16h prior stimulation in non-tissue cultured treated
TR:TREATMENT_SUMMARY             	6-well plate (BD Falcons). Cells were then stimulated with killed labelled
TR:TREATMENT_SUMMARY             	bacteria at MOI 25, centrifuged at 2000 rpm for 5 min. BMDMs were incubated for
TR:TREATMENT_SUMMARY             	5 min and washed with PBS to remove non-ingested bacteria and further incubated
TR:TREATMENT_SUMMARY             	for 4h.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Metabolites from frozen pellets were extracted at 4e6 cells per mL using ice
SP:SAMPLEPREP_SUMMARY            	cold 5:3:2 methanol:acetonitrile:water (v/v/v) with vigorous vortexing at 4
SP:SAMPLEPREP_SUMMARY            	degrees C followed by centrifugation as described for 10 min at 18,000 g.
SP:SAMPLEPREP_SUMMARY            	Supernatants were maintained at 4°C until analysis that same day.
SP:PROCESSING_STORAGE_CONDITIONS 	4℃
SP:EXTRACT_STORAGE               	-80℃
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Negative C18
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
CH:SOLVENT_A                     	95% water/5% acetonitrile; 1 mM ammonium acetate
CH:SOLVENT_B                     	95% acetonitrile/5% water; 1 mM ammonium acetate
CH:FLOW_GRADIENT                 	0-0.5 min 0% B, 0.5-1.1 min 0-100% B, 1.1-2.75 min hold at 100% B, 2.75-3 min
CH:FLOW_GRADIENT                 	100-0% B, 3-5 min hold at 0% B
CH:FLOW_RATE                     	450 uL/min
CH:COLUMN_TEMPERATURE            	45
CH:SAMPLE_INJECTION              	10 uL
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Orbitrap Exploris 120
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	We use a Thermo Orbitrap Exploris 120. Resolution 120,000, scan range 65-975
MS:MS_COMMENTS                   	m/z, maximum injection time 100 ms, microscans 1, automatic gain control (AGC)
MS:MS_COMMENTS                   	detection duration 20 msec, source voltage 2.0 kV, capillary temperature 320 C,
MS:MS_COMMENTS                   	vaporizer temp 200 C, and sheath gas 50, auxiliary gas 10, and sweep gas 1 (all
MS:MS_COMMENTS                   	nitrogen). Data converted to mzXML using RawConverter. Metabolites were
MS:MS_COMMENTS                   	annotated and integrated using Maven in conjunction with the KEGG database.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	peak area
MS_METABOLITE_DATA_START
Samples	AA89-33_r67+	AA89-34_r102+	AA89-35_r39+	AA89-36_r15+	AA89-37_r55+	AA89-38_r8+	AA89-39_r42+	AA89-40_r32+	AA89-41_r53+	AA89-42_r38+	AA89-43_r65+	AA89-44_r43+	AA89-45_r105+	AA89-46_r70+	AA89-47_r81+	AA89-48_r80+	AA89-49_r108+	AA89-50_r36+	AA89-51_r33+	AA89-52_r12+
Factors	treatment:CTRL | Sample source:murine bone marrow derived macrophages	treatment:CTRL | Sample source:murine bone marrow derived macrophages	treatment:CTRL | Sample source:murine bone marrow derived macrophages	treatment:CTRL | Sample source:murine bone marrow derived macrophages	treatment:Killed-Citro | Sample source:murine bone marrow derived macrophages	treatment:Killed-Citro | Sample source:murine bone marrow derived macrophages	treatment:Killed-Citro | Sample source:murine bone marrow derived macrophages	treatment:Killed-Citro | Sample source:murine bone marrow derived macrophages	treatment:Killed-Listeria | Sample source:murine bone marrow derived macrophages	treatment:Killed-Listeria | Sample source:murine bone marrow derived macrophages	treatment:Killed-Listeria | Sample source:murine bone marrow derived macrophages	treatment:Killed-Listeria | Sample source:murine bone marrow derived macrophages	treatment:Killed_Staph | Sample source:murine bone marrow derived macrophages	treatment:Killed_Staph | Sample source:murine bone marrow derived macrophages	treatment:Killed_Staph | Sample source:murine bone marrow derived macrophages	treatment:Killed_Staph | Sample source:murine bone marrow derived macrophages	treatment:Killed-Pseudo | Sample source:murine bone marrow derived macrophages	treatment:Killed-Pseudo | Sample source:murine bone marrow derived macrophages	treatment:Killed-Pseudo | Sample source:murine bone marrow derived macrophages	treatment:Killed-Pseudo | Sample source:murine bone marrow derived macrophages
Asparagine	220322	250348	263621	250090	52695	50628	51277	139726	293346	372525	365610	333018	37588	59755	54650	111124	168621	256524	158418	245867
Aspartate	2032964	2635771	2191431	2613602	2206826	1659573	2596073	2107713	3056631	2187642	2598002	3648387	1248929	763484	1446126	1831921	869129	758340	1810285	1578638
Glycine	365046	444479	312502	416590	346480	293900	337214	296521	313719	233344	272828	356400	138059	92657	178668	224874	140767	104081	281508	301458
Serine	355117	357549	303922	328212	324445	384648	385104	472385	375814	380382	442439	399486	269812	201870	191980	322954	190310	378502	262337	330798
Tyrosine	291639	337253	314839	210709	397129	463666	385036	289959	240129	307643	403501	216677	167138	229978	171840	220566	203290	184068	178335	183549
Cystine	33409	42077	25073	45316	15018	63248	34198	53977	28825	0	0	25749	15198	0	0	0	10570	0	21770	0
ADP	251396	460257	543700	429307	236691	152161	339652	563332	495557	402684	520467	648104	196657	251945	217954	137347	236747	259535	322004	194502
AMP	611018	557534	477131	411705	467293	524272	357227	159067	298139	297656	212387	696278	97863	98162	123137	123637	163132	118516	579243	356905
Adenosine	646888	720426	1213959	1175434	708615	911836	1017336	407331	1309538	318829	110711	962621	141566	125447	88565	249289	724672	343910	767443	383421
Adenine	2486744	1612331	2667060	1351493	2640667	1595104	1565081	855725	1634181	463510	706065	1062508	702383	205267	727290	711060	532585	543258	2317321	1156981
GDP	25220	63541	75412	136592	13794	19209	31796	125052	100763	67246	42919	125519	22816	67819	25512	40614	3423	26410	22106	26096
GMP	283218	437652	371072	347374	276770	436309	572649	495327	758631	497129	416156	505806	340871	128624	307206	360945	257419	200133	214892	325864
UDP	218943	366090	271034	376885	326299	315848	367980	473355	413688	388663	363900	567861	161889	203997	208669	216036	66563	188706	136683	239046
UMP	268593	471844	322711	329481	480198	567936	621205	661296	779747	537031	671646	434276	279870	291871	314899	268604	469433	407743	395767	278538
IDP	3366227	3691364	3523041	3874070	3401758	3032122	3949351	4127698	3223838	3020662	4329530	4188063	3732161	3266767	3917114	4276147	2922286	3707388	4683726	3538269
IMP	85574	108623	151195	55951	100675	102287	126905	156727	128886	190912	230283	154347	38085	24922	67791	26073	67188	107269	180651	201033
Inosine	3673161	4652166	4011699	2990721	1884195	2976481	2208737	2617620	4314268	3574322	2915882	2788111	814131	1161682	812376	730367	2114613	1524246	1513752	2012348
Hypoxanthine	208772	107659	153671	150886	141699	265021	97613	92672	213122	168676	223378	186904	113586	135144	232383	185248	217654	118966	235946	300282
Xanthine	607077	526677	270188	235480	361049	491011	193978	224616	525073	488679	312955	269634	105736	176780	144913	87280	310844	169354	124443	301968
Urate	391245	296166	553698	322573	65388	262022	243970	275911	1108575	1406601	888273	518188	221078	61775	232182	312213	233572	89152	365224	490864
UDP-glucose	397587	369807	354602	359448	830357	883286	779915	718642	460860	389721	303600	404653	148505	52609	64705	103947	270741	325592	239354	288338
ADP-D-ribose	27116	55840	22643	25496	81508	72348	22743	36706	57007	77421	59159	48273	0	0	0	0	30803	35583	56986	30834
Phosphate	125002464	116822048	142756080	136865664	115979208	113355000	139704864	138591584	148908544	147611072	167863696	144586304	192726192	240044416	189023376	174435968	207429328	192614352	136590592	162443184
Diphosphate	22185574	24821330	26604346	26497992	20959316	23226022	26723670	31247136	29997960	31843760	34242736	28084472	37339372	43159236	35555588	34881736	35275620	37780868	26575366	31382256
D-Glucose	274484	283360	1268758	362867	785521	801043	2068768	1518437	766854	749171	523889	627552	415324	303366	669638	647637	857437	684206	167188	320141
Hexose phosphate	233276	349990	321446	283506	443349	525147	659975	634371	540281	501688	465060	453724	211920	272377	265763	353225	294540	233338	351030	212135
D-Fructose 1-6-bisphosphate	31151	0	29936	20840	12838	20135	29283	66427	22981	40180	35359	19331	33363	37076	23427	38340	30773	0	0	0
D-Glyceraldehyde 3-phosphate/Glycerone phosphate	0	0	24468	14381	21353	17613	52437	52574	30028	25899	49471	0	54842	29085	90414	23523	58131	34929	45288	36134
2/3-Phospho-D-glycerate	332325	330876	333315	296283	1224440	1065288	1413750	1460617	649867	565928	670516	669846	522940	691887	742329	610266	253568	321786	320429	293022
Phosphoenolpyruvate	55193	96689	86473	85162	195146	229354	224325	231002	199174	238521	228484	125822	79780	50217	118457	59961	28022	99565	43965	36417
Pyruvate	853825	700536	763518	548540	846648	896158	591064	458297	526939	556007	452080	438268	424874	445340	498738	448488	694805	571060	678537	715205
Lactate	718550	464456	1554770	914205	658001	1156371	1174896	915384	752143	1018298	3404258	1523689	748429	312324	564700	523057	736028	585819	650402	950574
Maltose/Sucrose	75711	69186	55531	75618	188744	153210	254404	149923	85610	114665	147722	378463	42690	64527	112201	32560	117296	103539	43760	43247
Mannitol/Sorbitol/Glucitol/Iditol	86356	120610	428892	224352	150720	100002	533831	687236	1204821	1170697	1114492	834179	543146	438449	893841	798937	1495649	1112831	136456	430620
Ribose/Ribulose/Arabinose/Xylose/Xylulose	54184	70082	61084	17148	76245	17477	98787	73208	54157	41932	74120	25950	30104	19740	22610	11469	18175	15178	78333	33279
Arabitol/Xylitol/Ribitol	1079	8434	27592	25628	96722	45602	61021	38199	81810	81041	57504	68071	31060	34890	29166	36579	88509	70863	17433	40297
Citrate	2860382	3756881	4594616	4168432	18604098	23265482	26843306	28837656	5648376	4446548	4144836	4127447	32712950	45942804	43903596	48777024	3454711	2980884	1980449	2569374
2-Oxoglutarate	74113	86765	174037	62861	395493	454362	477973	436237	143322	148299	184340	163017	199993	246344	196679	163210	66310	62165	115695	91733
Succinate	315172	268898	340134	190584	5590088	6015681	6562324	6336302	1370293	1276211	1230127	1325053	627772	848186	683405	827417	824889	612494	562038	676951
Fumarate	127460	144773	166386	160914	160332	190652	217512	204403	299539	174902	223395	248363	85389	53329	113649	90830	116087	138336	96589	152624
Malate	1876268	1964026	2894222	2051647	1891872	2865622	2960108	3533482	4899074	3713426	3552847	4584961	1273182	1547982	1781545	1575847	2287373	2122105	2088913	2549858
2-Hydroxyglutarate/Citramalate	251467	295101	305204	294996	387618	446182	314415	424221	478492	507245	369300	423227	271583	469764	559064	471072	570417	355007	297903	279343
Itaconate	236615	277008	213744	271180	5256609	6560316	8063602	8681913	9926928	8581673	8003454	10166307	1834860	2112861	1501619	1583356	4612754	3657402	3681274	4690627
Sedoheptulose 1-phosphate	19619	18112	29704	17965	171359	194328	182980	163989	14462	19407	18555	21258	248916	267808	366202	447056	0	0	17951	24479
alpha-D-Ribose 1-phosphate	764269	834246	645280	793104	728103	900589	787694	909980	1062585	1024836	1135380	1027626	613993	623000	1089565	835686	923168	623406	797555	999682
5-Phospho-alpha-D-ribose 1-diphosphate	23282	29011	52560	28122	24120	39632	51251	22774	57088	53425	38489	47252	110515	97186	88898	80547	98559	130921	49536	50962
Glutathione (GSH)	1379930	2123559	1954081	1698374	1691099	2428554	4467373	3368606	5046975	3691551	4280007	2562894	1185730	302471	1228379	992060	1126855	1227841	1554266	1834704
Glutathione disulfide	181370	175769	234080	231106	674754	863216	731236	748324	156362	86923	68296	155618	78796	53613	124700	66425	37307	24929	46678	21196
5-Oxoproline	1836472	1211926	3784299	2775017	1653145	2527551	2998252	2047389	2142119	2865136	7846454	2600574	2590105	925107	1899501	1975230	1879596	1729060	1248148	2461224
S-Glutathionyl-L-cysteine	120846	186677	117640	112188	530445	390406	387676	496698	156619	189322	106545	84515	173661	121102	126225	79007	66979	27437	25649	36477
Ascorbate	34904	54983	33092	49299	33197	79936	63821	105768	49215	52056	49057	58380	75067	77066	61811	26044	46330	5755	73442	69058
Dehydroascorbate	109027	154512	157287	144533	489812	570940	588484	601257	186542	164284	125763	167084	756681	984854	792218	1073573	245398	124381	58178	127471
L-gamma-Glutamyl-L-hypoglycin	99809	296400	226425	205799	60546	89417	231050	513301	329765	255479	539340	358560	232506	255284	192285	239471	222642	271666	192148	239747
N-Acetylneuraminate	17486888	25869434	19909598	21409894	13614316	15344353	16793796	16071359	26361232	21756334	16285676	19960320	4109186	4770806	5377956	4652221	11979216	7787179	9254891	11311333
N-Glycoloyl-neuraminate	2447247	3813806	2644634	2802136	1726642	1901033	1769366	1991273	2963032	2575612	1722310	2346245	566349	488555	607168	604534	1526246	1032279	1235724	1677738
1-4-beta-D-Xylan	81590	95239	48520	111353	268886	312754	395751	374114	447598	309954	267869	371063	143786	191337	137980	159240	244141	131237	106184	161461
UDP-N-acetyl-D-glucosamine	526790	689812	626239	593558	426194	472184	435779	390586	761659	447344	348480	493017	164988	127256	165414	212694	348556	307906	421633	304368
CMP-N-acetylneuraminate	320315	494707	359015	394988	225578	204470	203067	228403	445667	391934	265284	411062	165576	51677	58426	123481	198649	212522	233951	237601
Phosphocreatine	121194	183431	210505	257452	46426	76729	72323	91456	176724	173608	162448	192039	0	26755	22818	0	55554	26269	26516	33884
Taurine	3324062	4432865	3901990	4322486	1386918	1815730	1719016	1259203	2867188	1931684	1455575	3146519	418092	256672	328526	429809	1314612	923723	1646032	1360910
Hypotaurine	33629	24070	27949	33998	15358	10504	19446	0	65978	44403	22229	49285	13902	0	0	0	13132	0	20004	24252
L-Cysteate	28904	23053	77479	26107	26517	98485	56949	45235	31967	98402	52721	36867	0	71297	24614	26347	123747	45497	42278	108020
Indole-3-acetaldehyde	134171	410994	230755	481931	426355	21420	418050	540051	99552	15797	338721	424010	61435	4431	75503	275799	1838	14785	188374	85764
N-formyl kynurenine	770391	863041	713977	645971	597456	681163	584211	353373	531955	387216	424395	786392	181163	187003	165773	145548	295439	267322	684887	355161
g-Oxalo-crotonate	373969	248952	474935	201982	358970	489974	417750	208669	546885	451934	356076	283465	396583	433131	518052	312625	632368	549505	573884	512785
2-Aminomuconate	220922	341340	242325	359780	378192	56521	299055	370504	136704	140853	313904	324949	162079	69778	101242	389107	129216	89488	314003	229084
Glycerol 3-phosphate	303805	316818	244964	282054	319314	287537	403515	386842	358445	316192	337361	309110	200869	127814	194122	171218	242357	206540	273789	329523
Ethanolamine phosphate	1714576	2284333	2165657	2337954	428163	847563	712167	837504	2194392	1756578	1160162	1611662	393519	209962	308055	212762	742252	355510	789888	818040
Sphingosine 1-phosphate	185373	217061	254269	209766	204527	132071	226051	291462	254107	185279	298287	292258	310721	136167	244917	316107	49425	177813	253677	226470
Hexanoic acid (caproate)	663533	743496	666083	597476	871580	891666	892181	704565	658722	610899	638999	700342	609894	586947	668963	746451	656862	745308	803103	812604
Octanoic acid (caprylate)	2748604	2914018	2735929	3080339	2929440	2987940	3020544	3023739	2811242	3048271	2877400	3345978	2733813	3132778	2881137	3344039	2344432	3118734	3018700	2782916
Nonanoic acid (pelargonate)	27334648	29750376	24477726	32039946	26139742	25108274	28059342	30138760	25103008	27108946	29064490	32999054	25792026	23993046	25345402	28304072	18681726	27743112	29626270	25425214
Decanoic acid (caprate)	30117274	36013708	30081728	39364400	37467364	29089518	36205516	34809728	29928170	29846330	31712646	35358992	28086546	28198880	30541920	35793064	19594054	28798290	32415866	28838496
Dodecanoic acid	10490565	13056748	9734224	15172804	11317793	11363819	12657567	13397485	9452343	10350601	10789585	15402720	10752767	10168057	9922000	12872949	9098405	9271226	11032028	10128117
Tetradecanoic acid	15065887	15888275	14328331	17751754	15990339	12348603	16028367	14457963	12282991	12036517	18626080	17466736	14831337	15339693	16265240	20292010	14810627	16272931	16130471	13854628
Hexadecanoic acid	152168672	191315024	208795952	235767040	163005520	171049072	234081744	214472576	170902928	179911568	226881104	223466560	196951744	183163568	166523824	220268464	140000176	187191088	207540400	199993024
Octadecanoic acid (stearic acid)	80948888	104208568	116089912	157108592	87039928	111038080	140537744	139732272	88494032	102241568	146061824	137571712	97339264	114336344	90538840	145838160	91257336	92130872	125682360	110693928
Tetradecenoic acid	444992	746386	538859	672909	836464	833254	870114	989386	613097	741336	1260990	912036	672604	664608	720987	911458	690904	537790	620052	581042
Octadecenoic acid (oleic acid)	33046766	40682588	39034860	45259136	40254964	51355332	50793660	54642416	49505972	47294688	55626932	52979940	50112624	50726804	44533212	55732700	39505452	45618916	46954692	49126900
Linoleate	3468872	4807872	4916161	5687576	4632431	7321817	9590504	7118136	6711048	6459782	9223731	8004482	6405111	7607901	6129974	9041039	4513054	5279710	7646031	7087544
Eicosatetraenoic acid	21387388	25598742	20702074	23367528	21677522	23627310	26598806	28390728	22389894	25727114	38749724	30005376	25867648	26534976	28516080	33663128	28040122	30416022	24220992	28773272
Eicosapentaenoic acid	3555910	4548486	3371485	4318303	3454735	4888950	4133562	5267709	6037904	7290739	7936992	7151518	5114403	6775878	5775173	6552397	5644698	6077402	5986078	7869338
Docosahexaenoic acid	13021068	17569486	15508711	18095050	15219061	17627600	18524590	21364770	21855466	21999836	26899386	22774578	18018002	21361126	18277244	22152290	17458042	20463306	19656786	21867766
(8Z-11Z-14Z)-Icosatrienoic acid	4162964	5773250	4302873	5692922	5061993	6332304	6156394	7471471	6598511	7169045	9270987	8134935	6049749	7060299	6157107	7211612	3897083	6389351	6165196	7174639
(7Z-10Z-13Z-16Z-19Z)-Docosa-7-10-13-16-19-pentaenoic acid	11022055	19520534	14830555	18612838	9635444	15048996	18456318	20063118	20054336	19681550	23564346	21354308	16493584	16653480	10446839	18768958	15656555	14615392	17123030	18837846
AMP 13C2	0	0	32846	16916	0	0	9076	45288	24099	49901	15229	0	25311	53676	0	16253	13093	17268	8349	30497
AMP 13C5	43159	34003	9776	0	31926	45422	27727	44632	24515	0	19523	18748	17679	43860	17717	19587	41523	22324	15649	18463
AMP 13C6	0	23555	0	12559	32212	19927	66691	22313	34122	19301	0	0	0	0	0	0	0	0	19922	20189
AMP 13C8	282597	327867	346821	301774	137405	190639	169958	96556	116470	56963	19216	241622	19879	0	0	34764	0	0	171162	96241
AMP 13C9	17941	19790	8359	20476	6146	0	8331	0	0	12386	0	21545	0	0	0	0	0	33287	6546	12648
AMP 13C10	0	0	0	0	35152	24523	11979	8170	0	0	0	18480	0	0	0	7968	0	0	9329	0
Adenosine 13C10	95359	104399	54443	78855	112403	63603	84646	98722	27729	18047	31339	50184	55165	38427	64952	41901	33864	19579	74559	65825
026 Adenine	2486744	1612331	2667060	1351493	2640667	1595104	1565081	855725	1634181	463510	706065	1062508	702383	205267	727290	711060	532585	543258	2317321	1156981
Adenine 13C5	0	0	0	0	119216	128019	55362	0	0	0	0	0	0	0	0	0	0	0	0	0
Citrate 13C2	7863	9798	55832	49022	207219	314368	394119	367362	73772	101278	101097	94060	318131	500053	486720	509690	66817	76656	24157	100317
Citrate 13C3	0	0	0	0	15609	54421	40826	111929	0	0	0	0	80691	162370	197634	173258	0	1047	0	0
Citrate 13C4	38182	9104	13131	18126	33721	43541	77443	131346	2536	25471	1891	1357	89220	124747	120659	172245	18111	1882	65704	49270
Citrate 13C6	16571	0	0	0	0	0	0	0	0	0	0	0	96215	74316	105825	85410	25833	0	0	0
Succinate 13C4	0	0	0	0	65118	32856	29571	0	0	0	0	0	0	0	0	0	0	0	0	0
Malate 13C2	9237	36992	12858	18568	22014	32726	43293	62748	53298	20143	78712	55207	5059	20478	23639	12325	24932	32306	25015	35450
Itaconate 13C2	0	0	0	0	6155	1829	34169	29824	62133	28402	32213	72979	0	0	0	0	16440	13300	41041	18532
2' 3'-Cyclic CMP 13C9	1173036	1116051	1501886	1054422	818589	1481496	1582638	1599272	1720086	1694357	1707049	1465019	1957164	2181666	1824875	1753755	1887136	2345346	1308940	1656327
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	KEGG ID	m/z	r.t.
Asparagine	C00152	131.046265	0.602
Aspartate	C00049	132.030258	0.565
Glycine	C00037	74.024742	0.607
Serine	C00065	104.035271	0.601
Tyrosine	C00082	180.066528	0.636
Cystine	C00491	239.016541	0.599
ADP	C00008	426.022003	0.558
AMP	C00020	346.055939	0.607
Adenosine	C00212	266.089264	0.637
Adenine	C00147	134.047287	0.638
GDP	C00035	442.017273	0.54
GMP	C00144	362.050751	0.588
UDP	C00015	402.99472	0.537
UMP	C00105	323.028717	0.582
IDP	C00104	427.007355	0.545
IMP	C00130	347.039734	0.589
Inosine	C00294	267.073242	0.621
Hypoxanthine	C00262	135.031265	0.682
Xanthine	C00385	151.026276	0.658
Urate	C00366	167.021118	0.601
UDP-glucose	C00029	565.047607	0.522
ADP-D-ribose	C01882	558.064331	0.552
Phosphate	C00009	96.96962	0.576
Diphosphate	C00013	176.935883	0.574
D-Glucose	C00031	179.055984	0.614
Hexose phosphate	C02965	259.022491	0.56
D-Fructose 1-6-bisphosphate	C00354	338.988739	0.533
D-Glyceraldehyde 3-phosphate/Glycerone phosphate	C00118	168.990784	0.565
2/3-Phospho-D-glycerate	C00631	184.985641	0.56
Phosphoenolpyruvate	C00074	166.975082	0.56
Pyruvate	C00022	87.008774	0.636
Lactate	C01432	89.024406	0.588
Maltose/Sucrose	C00208	341.109314	0.622
Mannitol/Sorbitol/Glucitol/Iditol	C00392	181.071762	0.617
Ribose/Ribulose/Arabinose/Xylose/Xylulose	C00121	149.045578	0.595
Arabitol/Xylitol/Ribitol	C01904	151.061462	0.606
Citrate	C00158	191.019608	0.543
2-Oxoglutarate	C00026	145.014267	0.554
Succinate	C00042	117.019432	0.559
Fumarate	C00122	115.003777	0.562
Malate	C00149	133.014221	0.549
2-Hydroxyglutarate/Citramalate	C02630	147.029907	0.559
Itaconate	C00490	129.019409	0.567
Sedoheptulose 1-phosphate	C06222	289.031738	0.576
alpha-D-Ribose 1-phosphate	C00620	229.01178	0.564
5-Phospho-alpha-D-ribose 1-diphosphate	C00119	388.93988	0.657
Glutathione (GSH)	C00051	306.077698	0.591
Glutathione disulfide	C00127	611.144592	0.546
5-Oxoproline	C01879	128.035263	0.588
S-Glutathionyl-L-cysteine	C05526	425.080475	0.578
Ascorbate	C00072	175.024734	0.57
Dehydroascorbate	C05422	173.009079	0.535
L-gamma-Glutamyl-L-hypoglycin	C08280	269.112091	0.634
N-Acetylneuraminate	C00270	308.098877	0.582
N-Glycoloyl-neuraminate	C03410	324.093719	0.574
1-4-beta-D-Xylan	C02352	131.035049	0.57
UDP-N-acetyl-D-glucosamine	C00043	606.07428	0.524
CMP-N-acetylneuraminate	C00128	613.140259	0.526
Phosphocreatine	C02305	210.028442	0.538
Taurine	C00245	124.007362	0.606
Hypotaurine	C00519	108.01252	0.611
L-Cysteate	C00506	167.997131	0.588
Indole-3-acetaldehyde	C00637	158.061218	1.948
N-formyl kynurenine	C02700	235.075821	0.611
g-Oxalo-crotonate	C03453	157.011871	0.661
2-Aminomuconate	C02220	156.029633	0.581
Glycerol 3-phosphate	C00093	171.006485	0.568
Ethanolamine phosphate	C00346	140.011749	0.589
Sphingosine 1-phosphate	C06124	378.240173	3.11
Hexanoic acid (caproate)	C01585	115.076538	1.786
Octanoic acid (caprylate)	C06423	143.107758	2.033
Nonanoic acid (pelargonate)	C01601	157.123413	2.12
Decanoic acid (caprate)	C01571	171.138992	2.204
Dodecanoic acid	C02679	199.170227	2.404
Tetradecanoic acid	C06424	227.201767	2.671
Hexadecanoic acid	C00249	255.232849	3.067
Octadecanoic acid (stearic acid)	C01530	283.264343	3.665
Tetradecenoic acid	C08322	225.186035	2.454
Octadecenoic acid (oleic acid)	C00712	281.248718	3.11
Linoleate	C01595	279.233093	2.778
Eicosatetraenoic acid	C00219	303.233093	2.679
Eicosapentaenoic acid	C06428	301.217438	2.502
Docosahexaenoic acid	C06429	327.233063	2.581
(8Z-11Z-14Z)-Icosatrienoic acid	C03242	305.248718	2.869
(7Z-10Z-13Z-16Z-19Z)-Docosa-7-10-13-16-19-pentaenoic acid	C16513	329.248718	2.708
AMP 13C2	C00020	348.062836	0.617
AMP 13C5	C00020	351.071838	0.621
AMP 13C6	C00020	352.081818	0.589
AMP 13C8	C00020	354.073364	0.601
AMP 13C9	C00020	355.081573	0.606
AMP 13C10	C00020	356.089355	0.605
Adenosine 13C10	C00212	276.11734	0.636
026 Adenine	c00047	134.047104	0.636
Adenine 13C5	c00047	139.064117	0.635
Citrate 13C2	C00158	193.025787	0.54
Citrate 13C3	C00158	194.029938	0.544
Citrate 13C4	C00158	195.031006	0.556
Citrate 13C6	C00158	197.037806	0.556
Succinate 13C4	C00042	121.033032	0.559
Malate 13C2	C00149	135.018967	0.552
Itaconate 13C2	C00490	131.025009	0.565
2' 3'-Cyclic CMP 13C9	C02354	313.065094	0.525
METABOLITES_END
#END