TY - JOUR
T1 - NMR analyses of the interaction between CCR5 and its ligand using functional reconstitution of CCR5 in lipid bilayers
AU - Yoshiura, Chie
AU - Kofuku, Yutaka
AU - Ueda, Takumi
AU - Mase, Yoko
AU - Yokogawa, Mariko
AU - Osawa, Masanori
AU - Terashima, Yuya
AU - Matsushima, Kouji
AU - Shimada, Ichio
PY - 2010/5/19
Y1 - 2010/5/19
N2 - CC-chemokine receptor 5 (CCR5) belongs to the G protein-coupled receptor (GPCR) family and plays important roles in the inflammatory response. In addition, its ligands inhibit the HIV infection. Structural analyses of CCR5 have been hampered by its instability in the detergent-solubilized form. Here, CCR5 was reconstituted into high density lipoprotein (rHDL), which enabled CCR5 to maintain its functions for >24 h and to be suitable for structural analyses. By applying the methyl-directed transferred cross-saturation (TCS) method to the complex between rHDL-reconstituted CCR5 and its ligand MIP-1α, we demonstrated that valine 59 and valine 63 of MIP-1α are in close proximity to CCR5 in the complex. Furthermore, these results suggest that the protective influence on HIV-1 infection of a SNP of MIP-1α is due to its change of affinity for CCR5. This method will be useful for investigating the various and complex signaling mediated by GPCR, and will also provide structural information about the interactions of other GPCRs with lipids, ligands, G-proteins, and effector molecules.
AB - CC-chemokine receptor 5 (CCR5) belongs to the G protein-coupled receptor (GPCR) family and plays important roles in the inflammatory response. In addition, its ligands inhibit the HIV infection. Structural analyses of CCR5 have been hampered by its instability in the detergent-solubilized form. Here, CCR5 was reconstituted into high density lipoprotein (rHDL), which enabled CCR5 to maintain its functions for >24 h and to be suitable for structural analyses. By applying the methyl-directed transferred cross-saturation (TCS) method to the complex between rHDL-reconstituted CCR5 and its ligand MIP-1α, we demonstrated that valine 59 and valine 63 of MIP-1α are in close proximity to CCR5 in the complex. Furthermore, these results suggest that the protective influence on HIV-1 infection of a SNP of MIP-1α is due to its change of affinity for CCR5. This method will be useful for investigating the various and complex signaling mediated by GPCR, and will also provide structural information about the interactions of other GPCRs with lipids, ligands, G-proteins, and effector molecules.
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U2 - 10.1021/ja100830f
DO - 10.1021/ja100830f
M3 - Article
C2 - 20423099
AN - SCOPUS:77952355281
SN - 0002-7863
VL - 132
SP - 6768
EP - 6777
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 19
ER -
