Grafting segments from the extracellular surface of CCR5 onto a bacteriorhodopsin transmembrane scaffold confers HIV-1 coreceptor activity.
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.
Abdulaev
Najmoutin G
NG
Center for Advanced Research in Biotechnology, National Institute of Standards and Technology and The University of Maryland Biotechnology Institute, Rockville, MD 20850, USA.
Strassmaier
Timothy T
TT
Ngo
Tony
T
Chen
Ruiwu
R
Luecke
Hartmut
H
Oprian
Daniel D
DD
Ridge
Kevin D
KD
D000595:Amino Acid Sequence; D000818:Animals; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019556:COS Cells; D002417:Cattle; D017930:Genes, Reporter; D015497:HIV-1 / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D019713:Receptors, CCR5 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015728:Receptors, HIV / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013057:Spectrum Analysis
D019713:Receptors, CCR5; D015728:Receptors, HIV; D011993:Recombinant Fusion Proteins; D001436:Bacteriorhodopsins