title: The griffithsin dimer is required for high-potency inhibition of HIV-1: evidence for manipulation of the structure of gp120 as part of the griffithsin dimer mechanism.
abstract: Griffithsin (Grft) is a protein lectin derived from red algae that tightly binds the HIV envelope protein gp120 and effectively inhibits virus infection. This inhibition is due to the binding by Grft of high-mannose saccharides on the surface of gp120. Grft has been shown to be a tight dimer, but the role of the dimer in Grft's anti-HIV function has not been fully explored. To investigate the role of the Grft dimer in anti-HIV function, an obligate dimer of Grft was designed by expressing the protein with a peptide linker between the two subunits. This "Grft-linker-Grft" is a folded protein dimer, apparently nearly identical in structural properties to the wild-type protein. A "one-armed" obligate dimer was also designed (Grft-linker-Grft OneArm), with each of the three carbohydrate binding sites of one subunit mutated while the other subunit remained intact. While both constructed dimers retained the ability to bind gp120 and the viral surface, Grft-linker-Grft OneArm was 84- to 1,010-fold less able to inhibit HIV than wild-type Grft, while Grft-linker-Grft had near-wild-type antiviral potency. Furthermore, while the wild-type protein demonstrated the ability to alter the structure of gp120 by exposing the CD4 binding site, Grft-linker-Grft OneArm largely lost this ability. In experiments to investigate gp120 shedding, it was found that Grft has different effects on gp120 shedding for strains from subtype B and subtype C, and this might correlate with Grft function. Evidence is provided that the dimer form of Grft is critical to the function of this protein in HIV inhibition.
authors.affiliation: University of California, Merced, Merced, California, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Xue
authors.forename: Bart
authors.initials: B
authors.lastname: Hoorelbeke
authors.forename: Ioannis
authors.initials: I
authors.lastname: Kagiampakis
authors.forename: Borries
authors.initials: B
authors.lastname: Demeler
authors.forename: Jan
authors.initials: J
authors.lastname: Balzarini
authors.forename: Patricia J
authors.initials: PJ
authors.lastname: Liwang
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology; D001665:Binding Sites; D015704:CD4 Antigens / Q000378:metabolism; D045744:Cell Line, Tumor; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015497:HIV-1 / Q000187:drug effects*; D006801:Humans; D037121:Plant Lectins / Q000494:pharmacology*; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D055503:Protein Multimerization; D015728:Receptors, HIV / Q000378:metabolism; D053586:Virus Internalization / Q000187:drug effects; D017201:Virus Shedding
chemical_list: D019380:Anti-HIV Agents; D015704:CD4 Antigens; D015699:HIV Envelope Protein gp120; D037121:Plant Lectins; D015728:Receptors, HIV; C499654:griffithsin protein, Griffithsia