NMR analyses of the interaction between CCR5 and its ligand using functional reconstitution of CCR5 in lipid bilayers

Chie Yoshiura, Yutaka Kofuku, Takumi Ueda, Yoko Mase, Mariko Yokogawa, Masanori Osawa, Yuya Terashima, Kouji Matsushima, Ichio Shimada

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57 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)6768-6777
Number of pages10
JournalJournal of the American Chemical Society
Volume132
Issue number19
DOIs
Publication statusPublished - 2010 May 19

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ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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