1. Introduction computer exercises Pre-processing LC-MS metabolomics
Antoine van Kampen
Bioinformatics laboratory
AMC

2. FAAH Fatty acid amide hydrolase or FAAH
Methoxy arachidonyl fluorophosphonate, commonly referred as MAFP, is an irreversible active site-directed enzyme inhibitor that inhibits nearly all serine hydrolases and serine proteases
Fatty acid amide hydrolase (FAAH) dimer shown with covalent inhibitor (MAFP, yellow) bound in the active site

3. FAAH (Fatty acid amide hydrolase)
FAAH is an integral membrane hydrolase with a single N-terminal transmembrane domain.
In vitro, FAAH has esterase and amidase activity.
In vivo, FAAH is the principal catabolic (breakdown) enzyme for a class of bioactive lipids called the fatty acid amides (FAAs).
Members of the FAAs include:
Anandamide (N-arachidonoylethanolamine), an endocannabinoid
Other N-acylethanolamines, such as N-oleoylethanolamine and N-palmitoylethanolamine
The sleep-inducing lipid oleamide
The N-acyltaurines, which are agonists of the transient receptor potential (TRP) family of calcium channels.
An esterase is a hydrolase enzyme that splits esters into an acid and an alcohol in a chemical reaction with water called hydrolysis.
Amidase catalyzes the hydrolysis of an amide (breaking of bonds by reaction with water).

4. Fatty Acid Amides
Fatty acid amides (FAAs) are amides formed from a fatty acid and an amine.
In biochemistry, FFAs which have ethanolamine as the amine component are known as N-acylethanolamines and they play an important role in chemical signalling
Chemical structure of oleamide, an FAA derived from the fatty acid oleic acid
wikipedia

5. Ethanol amide
From Piomelli D. (2003) The molecular logic of endocannabinoid signalling. Nature Reviews Neuroscience, 4(11), FAAH is part of endomembrane structures.

6. Untargeted metabolomics
Aim: identify substrate of FAAH in vivo
Approach: enzyme inactivation
Experiment: comparison of WT mice (FAAH(+/+)) to knock-out mice
(FAAH(-/-))
Mice: 3-6 months in age
6 WT / 6 KO
Tissue: brain and spinal cord
LC-MS

7. Total Ion Current
12 samples

8. One sample: WT 19

9. One sample: WT 19

10. Pre-processing steps during tutorial
FAAH
Knock-out
data