{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001441","ANALYSIS_ID":"AN002407","VERSION":"1","CREATED_ON":"August 6, 2020, 5:00 pm"},

"PROJECT":{"PROJECT_TITLE":"SUCLA2 mutations cause global protein succinylation contributing to the pathomechanism of a hereditary mitochondrial disease","PROJECT_SUMMARY":"Mitochondrial acyl-coenzyme A species are emerging as important sources of protein modification and damage. Succinyl-CoA ligase (SCL) deficiency causes a mitochondrial encephalomyopathy of unknown pathomechanism. Here, we show that succinyl-CoA accumulates in cells derived from patients carrying recessive mutations in the tricarboxylic acid cycle (TCA) gene succinyl-CoA ligase subunit beta (SUCLA2) causing global protein hyper-succinylation. Using mass spectrometry, we quantified nearly 1000 protein succinylation sites on 366 proteins from patient-derived fibroblasts and myotubes. Interestingly, hyper-succinylated proteins are distributed across cellular compartments, and many are known targets of the (NAD+)-dependent desuccinylase SIRT5. To test the contribution of hyper-succinylation to disease progression, we developed a zebrafish model of the SCL deficiency, and find that SIRT5 gain-of-function reduces global protein succinylation and improves survival. Thus, increased succinyl-CoA levels contribute to the pathology of SCL deficiency through post-translational modifications.","INSTITUTE":"North Carolina State University","LAST_NAME":"Liu","FIRST_NAME":"Xiaojing","ADDRESS":"Polk Hall, RM 128","EMAIL":"xliu68@ncsu.edu","PHONE":"9195154387"},

"STUDY":{"STUDY_TITLE":"Metabolomics of patient-derived fibroblasts","STUDY_SUMMARY":"7 control fibroblasts samples and 7 patient-derived fibroblasts samples were collected at day 0 and day 5. Intracellular metabolites were extracted from cells cultured in 6 well plate while acyl-CoA and 5-methyltetrahydrofolate were extracted from cells cultured in 60 mm dish.","INSTITUTE":"North Carolina State University","LAST_NAME":"Liu","FIRST_NAME":"Xiaojing","ADDRESS":"Polk Hall, RM 128","EMAIL":"xliu68@ncsu.edu","PHONE":"9195154387"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"D0-control1",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control1.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control2",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control2.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control3",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control3.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control4",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control4.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control5",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control5.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control6",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control6.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control7",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control7.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient1",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient1.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient2",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient2.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient3",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient3.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient4",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient4.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient5",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient5.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient6",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient6.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient7",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient7.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control1",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control1.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control2",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control2.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control3",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control3.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control4",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control4.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control5",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control5.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control6",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control6.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control7",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control7.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient1",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient1.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient2",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient2.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient3",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient3.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient4",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient4.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient5",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient5.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient6",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient6.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient7",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient7.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control1_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control1_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control2_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control2_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control3_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control3_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control4_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control4_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control5_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control5_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control6_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control6_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-control7_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-control7_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient1_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient1_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient2_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient2_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient3_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient3_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient4_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient4_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient5_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient5_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient6_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient6_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D0-patient7_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day0"},
"Additional sample data":{"RAW_FILE_NAME":"D0-patient7_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control1_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control1_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control2_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control2_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control3_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control3_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control4_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control4_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control5_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control5_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control6_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control6_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-control7_luna",
"Factors":{"Sample Source":"Control","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-control7_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient1_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient1_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient2_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient2_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient3_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient3_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient4_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient4_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient5_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient5_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient6_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient6_luna.raw"}
},
{
"Subject ID":"-",
"Sample ID":"D5-patient7_luna",
"Factors":{"Sample Source":"Patient","Treatment Duration":"Day5"},
"Additional sample data":{"RAW_FILE_NAME":"D5-patient7_luna.raw"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"7 control fibroblasts samples and 7 patient-derived fibroblasts samples were collected at day 0 and day 5. Intracellular metabolites were extracted from cells cultured in 6 well plate while acyl-CoA and 5-methyltetrahydrofolate were extracted from cells cultured in 60 mm dish. Intracellular metabolites and acyl-CoA from cells were harvested as described previously using 80% methanol/water as solvent (PMID: 24410464, PMID: 24894601, PMID: 25795660) and dry pellets were stored in -80 °C freezer until ready for LC-MS analysis. For acyl-CoA and 5-methyltetrahydrofolate analysis, dry pellets were reconstituted into 30 μL of sample solvent (water containing 50 mM ammonium acetate), and 10 μL was injected into the LC-MS. For non-acyl-CoA polar metabolite analysis, pellets were reconstituted into 30 μL of sample solvent (water:methanol:acetonitrile, 2:1:1, v/v), and 3 μL was injected into the LC-MS.","SAMPLE_TYPE":"Cultured fibroblasts"},

"TREATMENT":{"TREATMENT_SUMMARY":"Patient-derived fibroblasts cultured in standard culture conditions (proliferative condition) and cells cultured for 5 days in low-serum conditions (non-proliferative condition). Patients carry disease-causing mutations in SUCLA2. Controls are fibroblasts from age-matched patients with other mitochondrial diseases."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Intracellular metabolites and acyl-CoA from cells were harvested as described previously using 80% methanol/water as solvent (PMID: 24410464, PMID: 24894601, PMID: 25795660) and dry pellets were stored in -80 °C freezer until ready for LC-MS analysis. For acyl-CoA and 5-methyltetrahydrofolate analysis, dry pellets were reconstituted into 30 μL of sample solvent (water containing 50 mM ammonium acetate), and 10 μL was injected into the LC-MS. For non-acyl-CoA polar metabolite analysis, pellets were reconstituted into 30 μL of sample solvent (water:methanol:acetonitrile, 2:1:1, v/v), and 3 μL was injected into the LC-MS."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"HILIC method is for general metabolomics analysis.","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Waters Xbridge amide (100 × 2.1 mm i.d., 3.5 µm)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Exactive Plus Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"It was operated in the full-scan mode with the resolution set at 70 000 (at m/z 200).LC-MS data were analyzed using Sieve 2.0 (Thermo Scientific), and the integrated area under metabolite peak was used to compare the relative abundance of each metabolite in different samples in the same batch."},

"MS_METABOLITE_DATA":{
"Units":"peak area",

"Data":[{"Metabolite":"N-Methylethanolamine 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acid/Glycyl-glycine"},{"metabolite_name":"histamine"},{"metabolite_name":"acetamidopropanal"},{"metabolite_name":"Ornithine"},{"metabolite_name":"3-Ureidoisobutyrate/Glycylsarcosine"},{"metabolite_name":"C6H11NO3(4)"},{"metabolite_name":"(R)-mevalonate"},{"metabolite_name":"acetylcholine/4-(trimethylammonio)butanoate"},{"metabolite_name":"isoputreanine/putreanine"},{"metabolite_name":"Glycylproline/L-Prolinylglycine"},{"metabolite_name":"N-Acetylmethionine"},{"metabolite_name":"Pyridoxine/Norepinephrine-1"},{"metabolite_name":"Glycylleucine"},{"metabolite_name":"gama-L-glutamyl-L-alanine/5-L-Glutamyl-L-alanine-1"},{"metabolite_name":"C9H8O3(3)"},{"metabolite_name":"Adrenaline/L-Normetanephrine"},{"metabolite_name":"pseudoecgonine/ecgonine"},{"metabolite_name":"limonene/(+)-alpha-pinene"},{"metabolite_name":"indole-3-acetate/(5-hydroxyindol-3-yl)acetaldehyde"},{"metabolite_name":"(-)-Salsolinol/NN-dimethyldopaminequinone-1"},{"metabolite_name":"L-kynurenine"},{"metabolite_name":"methyl indole-3-acetate"},{"metabolite_name":"sphingosine/3-dehydrosphinganine"},{"metabolite_name":"3-oxo-10(R)-hydroxy-octadeca-6E8E12Z-trienoate/3-oxo-10(S)-hydroxy-octadeca-6E8E12Z-trienoate"},{"metabolite_name":"C18H32O4(3)"},{"metabolite_name":"C20H28O(5)"},{"metabolite_name":"C20H30O(4)"},{"metabolite_name":"C20H28O2(7)"},{"metabolite_name":"C27H44O2(5)"},{"metabolite_name":"C27H46O2(5)"},{"metabolite_name":"C27H44O3(5)"},{"metabolite_name":"C27H42O4(3)"},{"metabolite_name":"5beta-cholestane-3alpha-7alpha-12alpha-25-tetrol/5beta-cholestane-3alpha-7alpha-12alpha-26-tetrol"},{"metabolite_name":"5-guanidino-2-oxopentanoic acid"},{"metabolite_name":"phenylacetaldehyde"},{"metabolite_name":"L-serine"},{"metabolite_name":"L-cysteine"},{"metabolite_name":"NN-dimethylglycine"},{"metabolite_name":"L-asparagine"},{"metabolite_name":"L-homocysteine"},{"metabolite_name":"choline"},{"metabolite_name":"L-methionine"},{"metabolite_name":"L-histidine"},{"metabolite_name":"L-phenylalanine"},{"metabolite_name":"Biotin"},{"metabolite_name":"S-Adenosyl-L-homocysteine"},{"metabolite_name":"S-adenosyl-L-methionine"},{"metabolite_name":"Riboflavin"},{"metabolite_name":"Folate"},{"metabolite_name":"L-threonine"},{"metabolite_name":"glycine betaine"},{"metabolite_name":"L-glutamine"},{"metabolite_name":"L-tyrosine"},{"metabolite_name":"L-alanine"},{"metabolite_name":"L-proline"},{"metabolite_name":"adenosine"},{"metabolite_name":"2-deoxycytidine"},{"metabolite_name":"adenine"},{"metabolite_name":"Guanine"},{"metabolite_name":"cytosine"},{"metabolite_name":"thymine/imidazol-4-ylacetate"},{"metabolite_name":"(R)-carnitine"},{"metabolite_name":"O-acetylcarnitine"},{"metabolite_name":"propionyl-carnitine"},{"metabolite_name":"butyryl carnitine"},{"metabolite_name":"3-Hydroxybutyrylcarnitine"},{"metabolite_name":"succinyl carnitine"},{"metabolite_name":"tiglyl carnitine"},{"metabolite_name":"3-hydroxy-isovaleryl carnitine"},{"metabolite_name":"glutaryl carnitine"},{"metabolite_name":"octenoyl carnitine"},{"metabolite_name":"octanoyl carnitine/L-Octanoylcarnitine"},{"metabolite_name":"decenoyl carnitine"},{"metabolite_name":"decanoyl carnitine"},{"metabolite_name":"tetradecenoyl carnitine"},{"metabolite_name":"pendtadenoyl carnitine"},{"metabolite_name":"L-palmitoylcarnitine"},{"metabolite_name":"3-hydroxyhexadecenoylcarnitine"},{"metabolite_name":"3-hydroxyhexadecanoylcarnitine"},{"metabolite_name":"heptadecanoyl carnitine"},{"metabolite_name":"stearoylcarnitine"},{"metabolite_name":"tetradecanoyl carnitine(myristoyl carnitine)"},{"metabolite_name":"trans-Hexadec-2-enoyl carnitine"},{"metabolite_name":"alpha-linolenyl carnitine/gamma-linolenyl carnitine"},{"metabolite_name":"Linoleyl carnitine/Linoelaidyl carnitine"},{"metabolite_name":"Elaidic carnitine/Vaccenyl carnitine"},{"metabolite_name":"lauroyl carnitine"},{"metabolite_name":"1-methylpyrrolinium"},{"metabolite_name":"glycine"},{"metabolite_name":"sarcosine"},{"metabolite_name":"Trimethylamine N-oxide"},{"metabolite_name":"Methionine sulfoxide"}]
}

}