title: Nontargeted profiling of coenzyme A thioesters in biological samples by tandem mass spectrometry.
abstract: Coenzyme A (CoA) thioesters are ubiquitously present in metabolic networks and play a pivotal role in enzymatic formation and cleavage of carbon-carbon bonds. We present a method allowing nontargeted profiling of CoA-thioesters in biological samples. The reported UHPLC-MS/MS approach employes ion-pairing chromatography to separate CoA-metabolites carrying different chemical functionalities such as hydroxyl or multiple carboxyl groups and to distinguish between isomers. Selective detection of CoA-thioesters is accomplished by precursor ion scanning on a triple quadrupole mass spectrometer and takes advantage of the abundant fragment with m/z = -408 that originates from the CoA-moiety. We used a mixture of 19 commercially available CoA-derivatives to develop and optimize our method. As a proof of concept we demonstrated detection of the major CoA-intermediates of branched chain amino acid degradation in biological samples. We then applied our method to investigate degradation of lipids in the microorganism Mycobacterium smegmatis. Profiling of CoA-thioesters led to the discovery of a novel intermediate of cholesterol degradation. This demonstrates the power of our method for untargeted profiling of CoA-thioesters and adds a missing link in mycobacterial cholesterol catabolism.
authors.affiliation: Institute of Molecular Systems Biology, ETH Zurich, Wolfgang-Pauli Strasse 16, 8093 Zurich, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Zimmermann
authors.forename: Verena
authors.initials: V
authors.lastname: Thormann
authors.forename: Uwe
authors.initials: U
authors.lastname: Sauer
authors.forename: Nicola
authors.initials: N
authors.lastname: Zamboni
mesh_terms: D000818:Animals; D002417:Cattle; D003065:Coenzyme A / Q000032:analysis*; D004952:Esters / Q000032:analysis; D012710:Serum Albumin, Bovine / Q000032:analysis; D013438:Sulfhydryl Compounds / Q000032:analysis; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D004952:Esters; D013438:Sulfhydryl Compounds; D012710:Serum Albumin, Bovine; D003065:Coenzyme A