Lipid Discovery by Combinatorial Screening and Untargeted LC-MS/MS.
We present a method for the systematic identification of picogram quantities of new lipids in total extracts of tissues and fluids. It relies on the modularity of lipid structures and applies all-ions fragmentation LC-MS/MS and Arcadiate software to recognize individual modules originating from the same lipid precursor of known or assumed structure. In this way it alleviates the need to recognize and fragment very low abundant precursors of novel molecules in complex lipid extracts. In a single analysis of rat kidney extract the method identified 58 known and discovered 74 novel endogenous endocannabinoids and endocannabinoid-related molecules, including a novel class of N-acylaspartates that inhibit Hedgehog signaling while having no impact on endocannabinoid receptors.
Max Planck Institute for Cell Biology and Genetics, Pfotenhauerstraβe 108, 01307 Dresden, Germany.
Mesut
M
Bilgin
Max Planck Institute for Cell Biology and Genetics, Pfotenhauerstraβe 108, 01307 Dresden, Germany.
Petra
P
Born
Department of Experimental Medicine and Surgery, Tor Vergata University of Rome, via Montpellier 1, 00133, Rome, Italy.|European Center for Brain Research/Fondazione Santa Lucia, via del Fosso di Fiorano 65, 00143 Rome, Italy.
Filomena
F
Fezza
European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, Germany.
Michael
M
Heimes
Department of Medicine, Campus Bio-Medico University of Rome, Via Alvaro del Portillo 21, 00128 Rome.
Nicolina
N
Mastrangelo
Max Planck Institute for Cell Biology and Genetics, Pfotenhauerstraβe 108, 01307 Dresden, Germany.
Nicolai
N
Wagner
European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, Germany.
Carsten
C
Schultz
European Center for Brain Research/Fondazione Santa Lucia, via del Fosso di Fiorano 65, 00143 Rome, Italy.|Department of Medicine, Campus Bio-Medico University of Rome, Via Alvaro del Portillo 21, 00128 Rome.
Mauro
M
Maccarrone
Max Planck Institute for Cell Biology and Genetics, Pfotenhauerstraβe 108, 01307 Dresden, Germany.
Suzanne
S
Eaton
Max Planck Institute for Cell Biology and Genetics, Pfotenhauerstraβe 108, 01307 Dresden, Germany.|European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, Germany.
André
A
Nadler
Conway Institute of Biomolecular and Biomedical Research, University College Dublin, 4 Dublin, Ireland.
Matthias
M
Wilm
Max Planck Institute for Cell Biology and Genetics, Pfotenhauerstraβe 108, 01307 Dresden, Germany.
Andrej
A
Shevchenko
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D063388:Endocannabinoids / Q000032:analysis; D007668:Kidney / Q000737:chemistry; D008055:Lipids / Q000032:analysis*; D051381:Rats; D012984:Software; D053719:Tandem Mass Spectrometry / Q000379:methods*
D063388:Endocannabinoids; D008055:Lipids