TY - JOUR
T1 - Recombinant human PPAR-beta/delta ligand-binding domain is locked in an activated conformation by endogenous fatty acids
AU - Fyffe, Stewart A.
AU - Alphey, Magnus S.
AU - Buetow, Lori
AU - Smith, Terry K.
AU - Ferguson, Michael A. J.
AU - Sorensen, Morten D.
AU - Bjorkling, Fredrik
AU - Hunter, William N.
PY - 2006/3/3
Y1 - 2006/3/3
N2 - High-resolution crystallographic structures of recombinant human peroxisome proliferator-activated receptor ligand-binding domain (isotype beta/delta) reveal a fatty acid in the binding site. Mass spectrometry confirmed the presence of C16:0, C16:1, C18:0 and C18:1 in a ratio of approximately 3:2:1:4 with 11, Z-octadecenoic acid (cis-vaccenic acid) identified as the predominant species. These are endogenous fatty acids acquired from the bacterial expression system, and serve to lock the ligand-binding domain into the activated conformation. A requirement for crystal growth, the additive n-heptyl-beta-(D)-glucopyranoside, binds near the activation function helix where recognition of co-activator proteins occurs. Our observations suggest potential physiological ligands for human PPAR-beta/delta and highlight that reported binding studies must be treated with caution unless endogenous fatty acids have been removed from the sample prior to analysis. (c) 2005 Elsevier Ltd. All rights reserved.
AB - High-resolution crystallographic structures of recombinant human peroxisome proliferator-activated receptor ligand-binding domain (isotype beta/delta) reveal a fatty acid in the binding site. Mass spectrometry confirmed the presence of C16:0, C16:1, C18:0 and C18:1 in a ratio of approximately 3:2:1:4 with 11, Z-octadecenoic acid (cis-vaccenic acid) identified as the predominant species. These are endogenous fatty acids acquired from the bacterial expression system, and serve to lock the ligand-binding domain into the activated conformation. A requirement for crystal growth, the additive n-heptyl-beta-(D)-glucopyranoside, binds near the activation function helix where recognition of co-activator proteins occurs. Our observations suggest potential physiological ligands for human PPAR-beta/delta and highlight that reported binding studies must be treated with caution unless endogenous fatty acids have been removed from the sample prior to analysis. (c) 2005 Elsevier Ltd. All rights reserved.
U2 - 10.1016/j.jmb.2005.12.047
DO - 10.1016/j.jmb.2005.12.047
M3 - Article
C2 - 16405912
SN - 0022-2836
VL - 356
SP - 1005
EP - 1013
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 4
ER -