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A MBP-FAAH fusion protein as a tool to produce human and rat fatty acid amide hydrolase : expression and pharmacological comparison

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Labar, Geoffray [UCL] Vliet, Françoise V. Wouters, Johan [FUNDP] Lambert, Didier [UCL]

Fatty acid amide hydrolase (FAAH), a membrane-anchored enzyme responsible for the termination of endocannabinoid signalling, is an attractive target for treating conditions such as pain and anxiety. Inhibitors of the enzyme, optimized using rodent FAAH, are known but their pharmacology and medicinal chemistry properties on the human FAAH are missing. Therefore recombinant human enzyme would represent a powerful tool to evaluate new drug candidates. However, the production of high amounts of enzyme is hampered by the known refractiveness of FAAH to overexpression. Here, we report the successful overexpression of rat and human FAAH as a fusion to the E. coli maltose-binding protein, retaining catalytic properties of native FAAH. Several known FAAH inhibitors were tested and differences in their potencies toward the human and rat FAAH were found, underscoring the importance of using a human FAAH in the development of inhibitors.

Document type Article de périodique (Journal article) – Article de recherche – Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
Access type Accès restreint
Publication date 2008
Language Anglais
Journal information "Amino Acids" - Vol. 34, no. 1, p. 127-133 (2008)
Peer reviewed yes
Publisher Springer Wien ((Austria) Vienna)
issn 0939-4451
e-issn 1438-2199
Publication status Publié
Affiliation UCL - MD/FARM - Ecole de pharmacie
MESH Subject Amidohydrolases - antagonists & inhibitors - genetics - isolation & purification - metabolism ; Animals ; Carrier Proteins - genetics - isolation & purification - metabolism ; Enzyme Inhibitors - pharmacology ; Escherichia coli - genetics - metabolism ; Gene Expression ; Humans ; Molecular Weight ; Rats ; Recombinant Proteins - antagonists & inhibitors - genetics - isolation & purification - metabolism ; Solubility
Keywords Fatty acid amide hydrolase ; Endocannabinoid system ; Protein overexpression ; Fusion proteins ; Inhibitors characterization ; Maltose-binding protein
Links
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Bibliographic reference Labar, Geoffray ; Vliet, Françoise V. ; Wouters, Johan ; Lambert, Didier. A MBP-FAAH fusion protein as a tool to produce human and rat fatty acid amide hydrolase : expression and pharmacological comparison. In: Amino Acids, Vol. 34, no. 1, p. 127-133 (2008)
Permanent URL http://hdl.handle.net/2078.1/10953