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Multivalent interactions with gp120 are required for the anti-HIV activity of Cyanovirin.

ABSTRACT: Cyanovirin-N (CV-N) is a cyanobacterial lectin that binds to specific oligomannoses on the surface of gp120, resulting in nanomolar antiviral activity against HIV. In its monomeric form, CV-N contains two functional carbohydrate-binding domains, A and B. When refolded at high concentration, the protein can form a domain-swapped dimer. To clarify the role of multiple-binding sites in CV-N, we previously designed a monomeric mutant, P51G-m4-CVN, in which the binding site on domain A was rendered ineffective by four mutations (m4); in addition, a hinge region mutation (P51G) hinders the formation of a domain swapped dimer. The protein bound gp120 with diminished affinity and was completely inactive against HIV. Here, we present two mutants, DeltaQ50-m4-CVN and S52P-m4-CVN, which fold exclusively as domain-swapped dimers while containing the four mutations that abolish domain A. The dimers contain two intact B domains, thus restoring multivalency. DeltaQ50-m4-CVN and S52P-m4-CVN bind gp120 at low-nanomolar concentrations and recover in part the antiviral activity of wt CV-N. These results indicate that the number of carbohydrate binding domains, rather than their identity, is crucial to CV-N functionality.

SUBMITTER: Liu Y

PROVIDER: S-EPMC6961781 | biostudies-literature | 2009

REPOSITORIES: biostudies-literature

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Multivalent interactions with gp120 are required for the anti-HIV activity of Cyanovirin.

Liu Yinan Y Carroll Jacob R JR Holt Lindsey A LA McMahon James J Giomarelli Barbara B Ghirlanda Giovanna G

Biopolymers 20090101 3

Cyanovirin-N (CV-N) is a cyanobacterial lectin that binds to specific oligomannoses on the surface of gp120, resulting in nanomolar antiviral activity against HIV. In its monomeric form, CV-N contains two functional carbohydrate-binding domains, A and B. When refolded at high concentration, the protein can form a domain-swapped dimer. To clarify the role of multiple-binding sites in CV-N, we previously designed a monomeric mutant, P51G-m4-CVN, in which the binding site on domain A was rendered i ...[more]

PMID: 19235857

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Project description:Gut-homing ?4?7high CD4+ T lymphocytes have been shown to be preferentially targeted by human immunodeficiency virus type 1 (HIV-1) and are implicated in HIV-1 pathogenesis. Previous studies demonstrated that HIV-1 envelope protein gp120 binds and signals through ?4?7 and that this likely contributes to the infection of ?4?7high T cells and promotes cell-to-cell virus transmission. Structures within the second variable loop (V2) of gp120, including the tripeptide motif LDV/I, are thought to mediate gp120-?4?7 binding. However, lack of ?4?7 binding has been reported in gp120 proteins containing LDV/I, and the precise determinants of gp120-?4?7 binding are not fully defined. In this work, we report the novel finding that fibronectins mediate indirect gp120-?4?7 interactions. We show that Chinese hamster ovary (CHO) cells used to express recombinant gp120 produced fibronectins and other extracellular matrix proteins that copurified with gp120. CHO cell fibronectins were able to mediate the binding of a diverse panel of gp120 proteins to ?4?7 in an in vitro cell binding assay. The V2 loop was not required for fibronectin-mediated binding of gp120 to ?4?7, nor did V2-specific antibodies block this interaction. Removal of fibronectin through anion-exchange chromatography abrogated V2-independent gp120-?4?7 binding. Additionally, we showed a recombinant human fibronectin fragment mediated gp120-?4?7 interactions similarly to CHO cell fibronectin. These findings provide an explanation for the apparently contradictory observations regarding the gp120-?4?7 interaction and offer new insights into the potential role of fibronectin and other extracellular matrix proteins in HIV-1 biology.IMPORTANCE Immune tissues within the gut are severely damaged by HIV-1, and this plays an important role in the development of AIDS. Integrin ?4?7 plays a major role in the trafficking of lymphocytes, including CD4+ T cells, into gut lymphoid tissues. Previous reports indicate that some HIV-1 gp120 envelope proteins bind to and signal through ?4?7, which may help explain the preferential infection of gut CD4+ T cells. In this study, we demonstrate that extracellular matrix proteins can mediate interactions between gp120 and ?4?7 This suggests that the extracellular matrix may be an important mediator of HIV-1 interaction with ?4?7-expressing cells. These findings provide new insight into the nature of HIV-1-?4?7 interactions and how these interactions may represent targets for therapeutic intervention.

Dataset Information

Multivalent interactions with gp120 are required for the anti-HIV activity of Cyanovirin.

Publications

Multivalent interactions with gp120 are required for the anti-HIV activity of Cyanovirin.

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