{
"METABOLOMICS WORKBENCH":{"VERSION":"1","CREATED_ON":"September 20, 2017, 1:19 pm"},

"PROJECT":{"PROJECT_TITLE":"The ubiquitin ligase MuRF1 regulates PPARa activity in the heart by enhancing nuclear export via monoubiquitination","PROJECT_TYPE":"Targeted Metabolomics","PROJECT_SUMMARY":"The transcriptional regulation of peroxisome proliferator-activated receptor (PPAR) a by post-translational modification, such as ubiquitin, has not been described. We report here for the first time an ubiquitin ligase (muscle ring finger-1/MuRF1) that inhibits fatty acid oxidation by inhibiting PPARa, but not PPARß/d or PPAR? in cardiomyocytes in vitro. Similarly, MuRF1 Tg+ hearts showed significant decreases in nuclear PPARa activity and acyl-carnitine intermediates, while MuRF1-/- hearts exhibited increased PPARa activity and acyl-carnitine intermediates. MuRF1 directly interacts with PPARa, mono-ubiquitinates it, and targets it for nuclear export to inhibit fatty acid oxidation in a proteasome independent manner. We then identified a previously undescribed nuclear export sequence in PPARa, along with three specific lysines (292, 310, 388) required for MuRF1's targeting of nuclear export. These studies identify the role of ubiquitination in regulating cardiac PPARa, including the ubiquitin ligase that may be responsible for this critical regulation of cardiac metabolism in heart failure.","INSTITUTE":"University of North Carolina","DEPARTMENT":"McAllister Heart Institute, Department of Internal Medicine","LABORATORY":"Multiple Centers","LAST_NAME":"Willis","FIRST_NAME":"Monte","ADDRESS":"111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA","EMAIL":"monte_willis@med.unc.edu","PHONE":"919-360-7599","FUNDING_SOURCE":"NIH, Fondation Leducq, AHA mid-Atlantic affiliate, AHA scientist development grant, Jefferson-Pilot Corporation Fellowship in Academic Medicine"},

"STUDY":{"STUDY_TITLE":"Targeted metabolomics of MuRF1 overexpressing cardiomyocytes compared to their wildtype controls (part I)","STUDY_TYPE":"Targeted metabolomic analysis","STUDY_SUMMARY":"The transcriptional regulation of peroxisome proliferator-activated receptor (PPAR) a by post-translational modification, such as ubiquitin, has not been described. We report here for the first time an ubiquitin ligase (muscle ring finger-1/MuRF1) that inhibits fatty acid oxidation by inhibiting PPARa, but not PPARß/d or PPAR? in cardiomyocytes in vitro. Similarly, MuRF1 Tg+ hearts showed significant decreases in nuclear PPARa activity and acyl-carnitine intermediates, while MuRF1-/- hearts exhibited increased PPARa activity and acyl-carnitine intermediates. MuRF1 directly interacts with PPARa, mono-ubiquitinates it, and targets it for nuclear export to inhibit fatty acid oxidation in a proteasome independent manner. We then identified a previously undescribed nuclear export sequence in PPARa, along with three specific lysines (292, 310, 388) required for MuRF1's targeting of nuclear export. These studies identify the role of ubiquitination in regulating cardiac PPARa, including the ubiquitin ligase that may be responsible for this critical regulation of cardiac metabolism in heart failure.","INSTITUTE":"University of North Carolina","DEPARTMENT":"McAllister Heart Institute, Department of Internal Medicine","LABORATORY":"Multiple Centers","LAST_NAME":"Willis","FIRST_NAME":"Monte","ADDRESS":"111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA","EMAIL":"monte_willis@med.unc.edu","PHONE":"919-360-7599"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Heart 1",
"Factors":{"Genotype":"WT-MuRF1 tg+"}
},
{
"Subject ID":"-",
"Sample ID":"Heart 2",
"Factors":{"Genotype":"WT-MuRF1 tg+"}
},
{
"Subject ID":"-",
"Sample ID":"Heart 3",
"Factors":{"Genotype":"WT-MuRF1 tg+"}
},
{
"Subject ID":"-",
"Sample ID":"Heart 4",
"Factors":{"Genotype":"MuRF1 tg+"}
},
{
"Subject ID":"-",
"Sample ID":"Heart 5",
"Factors":{"Genotype":"MuRF1 tg+"}
},
{
"Subject ID":"-",
"Sample ID":"Heart 6",
"Factors":{"Genotype":"MuRF1 tg+"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"COS7 and H9C2 cells were transfected with PPRE-luc, pcDNA 3.1, ß-galactosidase, and the corresponding PPAR isoform (PPARa, PPARß/d or PPAR?) as indicated. 24 hours followings transfection cells were transduced with Ad.GFP-Myc-MuRF1. Cells were harvested 24hours later following observation of GFP by light microscopy"},

"TREATMENT":{"TREATMENT_SUMMARY":"N/A"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Acyl-carnitines were analyzed using stable isotope dilution techniques. Amino acids and acyl-carnitine measurements were made by flow injection tandem mass spectrometry"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"ACQUITY UPLC 1.7 µm column"},

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

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 5975","INSTRUMENT_TYPE":"Single quadrupole","MS_TYPE":"EI","ION_MODE":"POSITIVE"},

"MS_METABOLITE_DATA":{
"Units":"uM",

"Data":[{"Metabolite":"C2","Heart 1":"23.6000","Heart 2":"22.5000","Heart 3":"20.8000","Heart 4":"18.5000","Heart 5":"18.8000","Heart 6":"26.9000"},{"Metabolite":"C3","Heart 1":"0.3530","Heart 2":"0.9490","Heart 3":"0.6430","Heart 4":"0.8450","Heart 5":"0.6460","Heart 6":"0.9690"},{"Metabolite":"C4/Ci4","Heart 1":"0.0973","Heart 2":"0.0727","Heart 3":"0.0661","Heart 4":"0.0837","Heart 5":"0.0352","Heart 6":"0.0502"},{"Metabolite":"C5:1","Heart 1":"0.0137","Heart 2":"0.0412","Heart 3":"0.0505","Heart 4":"0.0367","Heart 5":"0.0443","Heart 6":"0.0436"},{"Metabolite":"C5's","Heart 1":"0.0294","Heart 2":"0.0257","Heart 3":"0.0447","Heart 4":"0.0365","Heart 5":"0.0279","Heart 6":"0.0248"},{"Metabolite":"C4-OH","Heart 1":"0.6950","Heart 2":"0.5340","Heart 3":"0.5530","Heart 4":"0.4500","Heart 5":"0.6190","Heart 6":"0.5900"},{"Metabolite":"C6","Heart 1":"0.1640","Heart 2":"0.0910","Heart 3":"0.0364","Heart 4":"0.0438","Heart 5":"0.0366","Heart 6":"0.0117"},{"Metabolite":"C5-OH/C3-DC","Heart 1":"0.5270","Heart 2":"0.6480","Heart 3":"0.9940","Heart 4":"0.8870","Heart 5":"0.5750","Heart 6":"0.7230"},{"Metabolite":"Ci4-DC/C4-DC","Heart 1":"0.3300","Heart 2":"0.3780","Heart 3":"0.3780","Heart 4":"0.4070","Heart 5":"0.2970","Heart 6":"0.3600"},{"Metabolite":"C8:1","Heart 1":"0.0041","Heart 2":"0.0170","Heart 3":"0.0105","Heart 4":"0.0105","Heart 5":"0.0111","Heart 6":"0.0079"},{"Metabolite":"C8","Heart 1":"0.0089","Heart 2":"0.0073","Heart 3":"0.0034","Heart 4":"0.0039","Heart 5":"0.0076","Heart 6":"0.0068"},{"Metabolite":"C5-DC","Heart 1":"0.0037","Heart 2":"0.0054","Heart 3":"0.0012","Heart 4":"0.0076","Heart 5":"0.0018","Heart 6":"0.0095"},{"Metabolite":"C6:1-DC/C8:1-OH","Heart 1":"0.0147","Heart 2":"0.0065","Heart 3":"0.0097","Heart 4":"0.0030","Heart 5":"0.0106","Heart 6":"0.0084"},{"Metabolite":"C6-DC","Heart 1":"0.0276","Heart 2":"0.0434","Heart 3":"0.0190","Heart 4":"0.0873","Heart 5":"0.0670","Heart 6":"0.0354"},{"Metabolite":"C10:3","Heart 1":"0.0012","Heart 2":"0.0005","Heart 3":"","Heart 4":"0.0013","Heart 5":"0.0033","Heart 6":"0.0016"},{"Metabolite":"C10:2","Heart 1":"0.0033","Heart 2":"0.0068","Heart 3":"0.0013","Heart 4":"0.0040","Heart 5":"0.0054","Heart 6":"0.0030"},{"Metabolite":"C10:1","Heart 1":"0.0117","Heart 2":"0.0115","Heart 3":"0.0059","Heart 4":"0.0070","Heart 5":"0.0059","Heart 6":"0.0069"},{"Metabolite":"C10","Heart 1":"0.0081","Heart 2":"0.0074","Heart 3":"0.0028","Heart 4":"0.0036","Heart 5":"","Heart 6":"0.0014"},{"Metabolite":"C7-DC","Heart 1":"0.0040","Heart 2":"0.0048","Heart 3":"0.0083","Heart 4":"0.0057","Heart 5":"0.0001","Heart 6":"0.0040"},{"Metabolite":"C8:1-DC","Heart 1":"0.0042","Heart 2":"","Heart 3":"0.0025","Heart 4":"0.0055","Heart 5":"0.0067","Heart 6":"0.0039"},{"Metabolite":"C10-OH/C8-DC","Heart 1":"0.0765","Heart 2":"0.1070","Heart 3":"0.0580","Heart 4":"0.0821","Heart 5":"0.1100","Heart 6":"0.0352"},{"Metabolite":"C12:1","Heart 1":"0.0063","Heart 2":"0.0068","Heart 3":"0.0025","Heart 4":"0.0035","Heart 5":"0.0010","Heart 6":""},{"Metabolite":"C12","Heart 1":"0.0081","Heart 2":"","Heart 3":"0.0007","Heart 4":"0.0006","Heart 5":"0.0026","Heart 6":"0.0011"},{"Metabolite":"C12-OH/C10-DC","Heart 1":"0.0759","Heart 2":"0.1140","Heart 3":"0.0584","Heart 4":"0.0765","Heart 5":"0.1070","Heart 6":"0.0331"},{"Metabolite":"C14:2","Heart 1":"0.0058","Heart 2":"0.0094","Heart 3":"0.0065","Heart 4":"0.0065","Heart 5":"0.0027","Heart 6":"0.0034"},{"Metabolite":"C14:1","Heart 1":"0.0117","Heart 2":"0.0098","Heart 3":"0.0056","Heart 4":"0.0098","Heart 5":"0.0051","Heart 6":"0.0033"},{"Metabolite":"C14","Heart 1":"0.0250","Heart 2":"0.0100","Heart 3":"0.0064","Heart 4":"0.0108","Heart 5":"0.0089","Heart 6":"0.0067"},{"Metabolite":"C14:1-OH/C12:1-DC","Heart 1":"0.1070","Heart 2":"0.1010","Heart 3":"0.0514","Heart 4":"0.0624","Heart 5":"0.0669","Heart 6":"0.0195"},{"Metabolite":"C14-OH/C12-DC","Heart 1":"0.0963","Heart 2":"0.0826","Heart 3":"0.0515","Heart 4":"0.0729","Heart 5":"0.1000","Heart 6":"0.0312"},{"Metabolite":"C16:2","Heart 1":"0.0145","Heart 2":"0.0058","Heart 3":"0.0036","Heart 4":"0.0052","Heart 5":"0.0042","Heart 6":"0.0029"},{"Metabolite":"C16:1","Heart 1":"0.0188","Heart 2":"0.0396","Heart 3":"0.0175","Heart 4":"0.0282","Heart 5":"0.0224","Heart 6":"0.0099"},{"Metabolite":"C16","Heart 1":"0.0711","Heart 2":"0.0295","Heart 3":"0.0215","Heart 4":"0.0189","Heart 5":"0.0173","Heart 6":"0.0125"},{"Metabolite":"C16:1-OH/C14:1-DC","Heart 1":"0.0748","Heart 2":"0.0839","Heart 3":"0.0424","Heart 4":"0.0351","Heart 5":"0.0482","Heart 6":"0.0109"},{"Metabolite":"C16-OH/C14-DC","Heart 1":"0.1230","Heart 2":"0.1020","Heart 3":"0.0375","Heart 4":"0.0457","Heart 5":"0.0652","Heart 6":"0.0198"},{"Metabolite":"C18:2","Heart 1":"0.0357","Heart 2":"0.0555","Heart 3":"0.0391","Heart 4":"0.0301","Heart 5":"0.0403","Heart 6":"0.0119"},{"Metabolite":"C18:1","Heart 1":"0.0403","Heart 2":"0.0456","Heart 3":"0.0191","Heart 4":"0.0330","Heart 5":"0.0357","Heart 6":"0.0224"},{"Metabolite":"C18","Heart 1":"0.0431","Heart 2":"0.0192","Heart 3":"0.0154","Heart 4":"0.0130","Heart 5":"0.0071","Heart 6":"0.0080"},{"Metabolite":"C18:2-OH","Heart 1":"0.1010","Heart 2":"0.0536","Heart 3":"0.0235","Heart 4":"0.0190","Heart 5":"0.0318","Heart 6":"0.0044"},{"Metabolite":"C18:1-OH/C16:1-DC","Heart 1":"0.1340","Heart 2":"0.0987","Heart 3":"0.0484","Heart 4":"0.0434","Heart 5":"0.0625","Heart 6":"0.0183"},{"Metabolite":"C18-OH/C16-DC","Heart 1":"0.0606","Heart 2":"0.0466","Heart 3":"0.0249","Heart 4":"0.0295","Heart 5":"0.0386","Heart 6":"0.0131"},{"Metabolite":"C20:4","Heart 1":"0.0048","Heart 2":"0.0104","Heart 3":"0.0064","Heart 4":"0.0154","Heart 5":"0.0067","Heart 6":"0.0047"},{"Metabolite":"C20","Heart 1":"0.0138","Heart 2":"0.0054","Heart 3":"0.0025","Heart 4":"0.0025","Heart 5":"0.0042","Heart 6":"0.0018"},{"Metabolite":"C20:1-OH/C18:1-DC","Heart 1":"0.0169","Heart 2":"0.0189","Heart 3":"0.0080","Heart 4":"0.0094","Heart 5":"0.0170","Heart 6":"0.0140"},{"Metabolite":"C20-OH/C18-DC","Heart 1":"0.0067","Heart 2":"0.0069","Heart 3":"0.0033","Heart 4":"0.0037","Heart 5":"0.0048","Heart 6":"0.0037"},{"Metabolite":"C22","Heart 1":"0.0046","Heart 2":"","Heart 3":"","Heart 4":"0.0027","Heart 5":"0.0030","Heart 6":"0.0011"}],

"Metabolites":[{"metabolite_name":"C2"},{"metabolite_name":"C3"},{"metabolite_name":"C4/Ci4"},{"metabolite_name":"C5:1"},{"metabolite_name":"C5's"},{"metabolite_name":"C4-OH"},{"metabolite_name":"C6"},{"metabolite_name":"C5-OH/C3-DC"},{"metabolite_name":"Ci4-DC/C4-DC"},{"metabolite_name":"C8:1"},{"metabolite_name":"C8"},{"metabolite_name":"C5-DC"},{"metabolite_name":"C6:1-DC/C8:1-OH"},{"metabolite_name":"C6-DC"},{"metabolite_name":"C10:3"},{"metabolite_name":"C10:2"},{"metabolite_name":"C10:1"},{"metabolite_name":"C10"},{"metabolite_name":"C7-DC"},{"metabolite_name":"C8:1-DC"},{"metabolite_name":"C10-OH/C8-DC"},{"metabolite_name":"C12:1"},{"metabolite_name":"C12"},{"metabolite_name":"C12-OH/C10-DC"},{"metabolite_name":"C14:2"},{"metabolite_name":"C14:1"},{"metabolite_name":"C14"},{"metabolite_name":"C14:1-OH/C12:1-DC"},{"metabolite_name":"C14-OH/C12-DC"},{"metabolite_name":"C16:2"},{"metabolite_name":"C16:1"},{"metabolite_name":"C16"},{"metabolite_name":"C16:1-OH/C14:1-DC"},{"metabolite_name":"C16-OH/C14-DC"},{"metabolite_name":"C18:2"},{"metabolite_name":"C18:1"},{"metabolite_name":"C18"},{"metabolite_name":"C18:2-OH"},{"metabolite_name":"C18:1-OH/C16:1-DC"},{"metabolite_name":"C18-OH/C16-DC"},{"metabolite_name":"C20:4"},{"metabolite_name":"C20"},{"metabolite_name":"C20:1-OH/C18:1-DC"},{"metabolite_name":"C20-OH/C18-DC"},{"metabolite_name":"C22"}]
}

}