Project 6: Mothes/Blanchard Conformational trajectories of inhibitor-bound HIV-1 envelope Summary The design of effective antiviral therapies directed against HIV Env pursued by this program requires a fast method to determine the underlying molecular mechanism of HIV Env inhibition by small molecule inhibitors. Towards this end, we have established single-molecule Fluorescence Resonance Energy Transfer (smFRET) methods to visualize directly the structural dynamics of HIV Env in the context of the complete trimer on the surface of HIV virions. As a proof of concept we have successfully applied smFRET to the X4-tropic HIV-1 NL4-3- Our work reveals that the unliganded HIV Env is conformationally dynamic and intrinsically capable of sampling receptor CD4- and 17b-stabilized conformations. Importantly, the co-receptor binding site is more likely to be accessed from the CD4-induced state. Binding of CD4 lowers the energy of the CD4-stabilized conformation and prepares the HIV Env for co-receptor binding. These data explain how HIV Env is activated in a stepwise manner by CD4 and the co-receptor. Here, we will extend the application of smFRET to include physiologically relevant R5-tropic HIV-1 Env isolates. Dually modified HIV-1 Env variants will be used to determine the conformational trajectories of HIV Env induced by CD4 mimetics and antagonists. This will allow distinction between CD4 mimics such as NBD556 that induce CD4-like conformational changes and antagonists such as DMJ-l-228. smFRET imaging will also permit an understanding of conformational molecular events underlying HlV-1 inactivation either by prematurely activating Env, kinetic traps or allosterically diverting Env into off-pathways. As such, these smFRET technologies will provide a critical tool to determine the underlying molecular mechanism of HIV Env antagonism and inactivation developed by the Program Project Team and aid in a rational design of antiviral therapies that exploit the intrinsic vulnerabilities of HIV. smFRET technologies will accelerate pharmaceutical discovery by enabling screening for desired conformational effects that block, trap or inactivate HIV-1 Env.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Program Projects (P01)
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Special Emphasis Panel (ZRG1-AARR-E)
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Drexel University
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Clarkson, Benjamin R; Schön, Arne; Freire, Ernesto (2016) Conformational stability and self-association equilibrium in biologics. Drug Discov Today 21:342-7
Moraca, Francesca; Acharya, Kriti; Melillo, Bruno et al. (2016) Computational Evaluation of HIV-1 gp120 Conformations of Soluble Trimeric gp140 Structures as Targets for de Novo Docking of First- and Second-Generation Small-Molecule CD4 Mimics. J Chem Inf Model 56:2069-2079
Melillo, Bruno; Liang, Shuaiyi; Park, Jongwoo et al. (2016) Small-Molecule CD4-Mimics: Structure-Based Optimization of HIV-1 Entry Inhibition. ACS Med Chem Lett 7:330-4
Schön, Arne; Freire, Ernesto (2016) Enthalpy screen of drug candidates. Anal Biochem 513:1-6
Pustylnikov, Sergey; Dave, Rajnish S; Khan, Zafar K et al. (2016) Short Communication: Inhibition of DC-SIGN-Mediated HIV-1 Infection by Complementary Actions of Dendritic Cell Receptor Antagonists and Env-Targeting Virus Inactivators. AIDS Res Hum Retroviruses 32:93-100
Lee, Wen Shi; Richard, Jonathan; Lichtfuss, Marit et al. (2016) Antibody-Dependent Cellular Cytotoxicity against Reactivated HIV-1-Infected Cells. J Virol 90:2021-30
Madani, Navid; Princiotto, Amy M; Easterhoff, David et al. (2016) Antibodies Elicited by Multiple Envelope Glycoprotein Immunogens in Primates Neutralize Primary Human Immunodeficiency Viruses (HIV-1) Sensitized by CD4-Mimetic Compounds. J Virol 90:5031-46
Kalyana Sundaram, Ramalingam Venkat; Li, Huiyuan; Bailey, Lauren et al. (2016) Impact of HIV-1 Membrane Cholesterol on Cell-Independent Lytic Inactivation and Cellular Infectivity. Biochemistry 55:447-58
Schön, Arne; Freire, Ernesto (2016) Three easy pieces. Biochim Biophys Acta 1860:975-80
Herschhorn, Alon; Ma, Xiaochu; Gu, Christopher et al. (2016) Release of gp120 Restraints Leads to an Entry-Competent Intermediate State of the HIV-1 Envelope Glycoproteins. MBio 7:

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