Grafting Segments from the Extracellular Surface of CCR5 onto a Bacteriorhodopsin Transmembrane Scaffold Confers HIV-1 Coreceptor Activity

• Published in: Structure (London) Vol. 10; no. 4; pp. 515 - 525
• Main Authors: Abdulaev, Najmoutin G, Strassmaier, Timothy T, Ngo, Tony, Chen, Ruiwu, Luecke, Hartmut, Oprian, Daniel D, Ridge, Kevin D
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.04.2002
• Subjects: Amino Acid Sequence; Animals; bacteriorhodopsin; Bacteriorhodopsins - chemistry; Bacteriorhodopsins - genetics; Bacteriorhodopsins - metabolism; Cattle; CCR5; chimera; COS Cells; G protein-coupled receptor; Genes, Reporter; HIV-1; HIV-1 - chemistry; HIV-1 - metabolism; Humans; membrane protein folding; Molecular Sequence Data; Protein Structure, Secondary; Receptors, CCR5 - chemistry; Receptors, CCR5 - genetics; Receptors, CCR5 - metabolism; Receptors, HIV - chemistry; Receptors, HIV - metabolism; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Spectrum Analysis; chimera; G protein-coupled receptor; HIV-1; CCR5; bacteriorhodopsin; membrane protein folding
• ISSN: 0969-2126; 1878-4186
• DOI: 10.1016/S0969-2126(02)00752-9

Components from the extracellular surface of CCR5 interact with certain macrophage-tropic strains of human immunodeficiency virus type 1 (HIV-1) to mediate viral fusion and entry. To mimic these viral interacting site(s), the amino-terminal and extracellular loop segments of CCR5 were linked in tandem to form concatenated polypeptides, or grafted onto a seven-transmembrane bacteriorhodopsin scaffold to generate several chimeras. The chimera studies identified specific regions in CCR5 that confer HIV-1 coreceptor function, structural rearrangements in the transmembrane region that may modulate this activity, and a role for the extracellular surface in folding and assembly. Methods developed here may be applicable to the dissection of functional domains from other seven-transmembrane receptors and form a basis for future structural studies.