TY - JOUR
T1 - A MBP-FAAH fusion protein as a tool to produce human and rat fatty acid amide hydrolase
T2 - Expression and pharmacological comparison
AU - Labar, Geoffray
AU - Vliet, F. V.
AU - Wouters, Johan
AU - Lambert, D. M.
PY - 2008/1/1
Y1 - 2008/1/1
N2 - 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.
AB - 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.
KW - Endocannabinoid system
KW - Fatty acid amide hydrolase
KW - Fusion proteins
KW - Inhibitors characterization
KW - Maltose-binding protein
KW - Protein overexpression
UR - http://www.scopus.com/inward/record.url?scp=38149138066&partnerID=8YFLogxK
U2 - 10.1007/s00726-007-0540-1
DO - 10.1007/s00726-007-0540-1
M3 - Article
C2 - 17476568
AN - SCOPUS:38149138066
SN - 0939-4451
VL - 34
SP - 127
EP - 133
JO - Amino acids
JF - Amino acids
IS - 1
ER -