1. Conformational Stabilization of Gp41-Mimetic Miniproteins Opens Up New Ways of Inhibiting HIV-1 Fusion
- Author
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Mario Cano-Muñoz, Julie Lucas, Li-Yun Lin, Samuele Cesaro, Christiane Moog, and Francisco Conejero-Lara
- Subjects
N-terminal domain ,fusion inhibitor ,calorimetry ,coiled-coil ,envelope glycoprotein ,antiviral therapy ,gp41 ,Fusion inhibitors ,Protein Conformation ,viruses ,Organic Chemistry ,Envelope glycoprotein ,General Medicine ,Calorimetry ,Antiviral therapy ,Catalysis ,HIV Envelope Protein gp41 ,Computer Science Applications ,Inorganic Chemistry ,HIV Fusion Inhibitors ,HIV-1 ,Coiled-coil ,Amino Acid Sequence ,Disulfides ,Physical and Theoretical Chemistry ,Molecular Biology ,Spectroscopy - Abstract
Supplementary Materials: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/ijms23052794/s1., Acknowledgments: M.C.-M. acknowledges a grant from Youth Employment Operative Program of the Andalusia Government and the European Social Fund (ESF). S.C. acknowledges an exchange studentship from the ERASMUS+ program of the European Union. The results shown are included as part of M.C.-M. doctoral thesis., Inhibition of the HIV-1 fusion process constitutes a promising strategy to neutralize the virus at an early stage before it enters the cell. In this process, the envelope glycoprotein (Env) plays a central role by promoting membrane fusion. We previously identified a vulnerability at the flexible C-terminal end of the gp41 C-terminal heptad repeat (CHR) region to inhibition by a single-chain miniprotein (named covNHR-N) that mimics the first half of the gp41 N-terminal heptad repeat (NHR). The miniprotein exhibited low stability, moderate binding to its complementary CHR region, both as an isolated peptide and in native trimeric Envs, and low inhibitory activity against a panel of pseudoviruses. The addition of a disulfide bond stabilizing the miniprotein increased its inhibitory activity, without altering the binding affinity. Here, to further study the effect of conformational stability on binding and inhibitory potency, we additionally stabilized these miniproteins by engineering a second disulfide bond stapling their N-terminal end, The new disulfide-bond strongly stabilizes the protein, increases binding affinity for the CHR target and strongly improves inhibitory activity against several HIV-1 strains. Moreover, high inhibitory activity could be achieved without targeting the preserved hydrophobic pocket motif of gp41. These results may have implications in the discovery of new strategies to inhibit HIV targeting the gp41 CHR region., Grants BIO2016-76640-R and PID2019.107515RB.C21 from the Spain’s State Research Agency, SRA/10.13039/501100011033, co-funded by ERDF/ESF, “A way to make Europe”/“Investing in your future”
- Published
- 2022