A Cell Free Biomembrane Platform for Multimodal Study of Influenza Virus Hemagglutinin and for Evaluation of Entry-inhibitors Against Hemagglutinin

Overview

Journal Front Mol Biosci

Specialty Biology

Date 2022 Oct 31

PMID 36310596

Authors

Arpita Roy

Sylvester Byrne

Nirod Kumar Sarangi

Paul V Murphy

Tia E Keyes

Affiliations

Soon will be listed here.

Abstract

Seasonal periodic pandemics and epidemics caused by Influenza A viruses (IAVs) are associated with high morbidity and mortality worldwide. They are frequent and unpredictable in severity so there is a need for biophysical platforms that can be used to provide both mechanistic insights into influenza virulence and its potential treatment by anti-IAV agents. Host membrane viral association through the glycoprotein hemagglutinin (HA) of IAVs is one of the primary steps in infection. HA is thus a potential target for drug discovery and development against influenza. Deconvolution of the multivalent interactions of HA at the interfaces of the host cell membrane can help unravel therapeutic targets. In this contribution, we reported the effect of a multivalent HA glycoprotein association on various glycosphingolipid receptors (GD1a, GM3, GM1) doped asymmetrically into an artificial host membrane spanned across an aqueous filled microcavity array. The extent of HA association and its impact on membrane resistance, capacitance, and diffusivity was measured using highly sensitive electrochemical impedance spectroscopy (EIS) and fluorescence lifetime correlation spectroscopy (FLCS). Furthermore, we investigated the inhibition of the influenza HA glycoprotein association with the host mimetic surface by natural and synthetic sialic acid-based inhibitors (sialic acid, Siaα2,3-GalOMe, FB127, 3-sialyl lactose) using electrochemical impedance spectroscopy and observe that while all inhibit, they do not prevent host binding. Overall, the work demonstrates the platform provides a label-free screening platform for the biophysical evaluation of new inhibitors in the development of potential therapeutics for IAV infection prevention and treatment.

Citing Articles

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References

Srinivasan A, Viswanathan K, Raman R, Chandrasekaran A, Raguram S, Tumpey T . Quantitative biochemical rationale for differences in transmissibility of 1918 pandemic influenza A viruses. Proc Natl Acad Sci U S A. 2008; 105(8):2800-5. PMC: 2268540. DOI: 10.1073/pnas.0711963105. View

Parton D, Tek A, Baaden M, Sansom M . Formation of raft-like assemblies within clusters of influenza hemagglutinin observed by MD simulations. PLoS Comput Biol. 2013; 9(4):e1003034. PMC: 3623702. DOI: 10.1371/journal.pcbi.1003034. View

Shi J, Yang T, Kataoka S, Zhang Y, Diaz A, Cremer P . GM1 clustering inhibits cholera toxin binding in supported phospholipid membranes. J Am Chem Soc. 2007; 129(18):5954-61. PMC: 3475181. DOI: 10.1021/ja069375w. View

Tang T, Savva A, Traberg W, Xu C, Thiburce Q, Liu H . Functional Infectious Nanoparticle Detector: Finding Viruses by Detecting Their Host Entry Functions Using Organic Bioelectronic Devices. ACS Nano. 2021; 15(11):18142-18152. DOI: 10.1021/acsnano.1c06813. View

Shkirskiy V, Kang M, McPherson I, Bentley C, Wahab O, Daviddi E . Electrochemical Impedance Measurements in Scanning Ion Conductance Microscopy. Anal Chem. 2020; 92(18):12509-12517. DOI: 10.1021/acs.analchem.0c02358. View

Li Y, Liu D, Wang Y, Su W, Liu G, Dong W . The Importance of Glycans of Viral and Host Proteins in Enveloped Virus Infection. Front Immunol. 2021; 12:638573. PMC: 8116741. DOI: 10.3389/fimmu.2021.638573. View

Hess S, Gould T, Gudheti M, Maas S, Mills K, Zimmerberg J . Dynamic clustered distribution of hemagglutinin resolved at 40 nm in living cell membranes discriminates between raft theories. Proc Natl Acad Sci U S A. 2007; 104(44):17370-5. PMC: 2077263. DOI: 10.1073/pnas.0708066104. View

Lam A, Kirkland O, Anderson P, Seetharaman N, Vujovic D, Thibault P . Single-virus assay reveals membrane determinants and mechanistic features of Sendai virus binding. Biophys J. 2022; 121(6):956-965. PMC: 8943810. DOI: 10.1016/j.bpj.2022.02.011. View

Gaus K, Gratton E, Kable E, Jones A, Gelissen I, Kritharides L . Visualizing lipid structure and raft domains in living cells with two-photon microscopy. Proc Natl Acad Sci U S A. 2003; 100(26):15554-9. PMC: 307606. DOI: 10.1073/pnas.2534386100. View

10.

Sauter N, Hanson J, Glick G, Brown J, Crowther R, Park S . Binding of influenza virus hemagglutinin to analogs of its cell-surface receptor, sialic acid: analysis by proton nuclear magnetic resonance spectroscopy and X-ray crystallography. Biochemistry. 1992; 31(40):9609-21. DOI: 10.1021/bi00155a013. View

11.

Siontorou C, Nikoleli G, Nikolelis D, Karapetis S . Artificial Lipid Membranes: Past, Present, and Future. Membranes (Basel). 2017; 7(3). PMC: 5618123. DOI: 10.3390/membranes7030038. View

12.

Du R, Cui Q, Rong L . Competitive Cooperation of Hemagglutinin and Neuraminidase during Influenza A Virus Entry. Viruses. 2019; 11(5). PMC: 6563287. DOI: 10.3390/v11050458. View

13.

Verma D, Gupta D, Lal S . Host Lipid Rafts Play a Major Role in Binding and Endocytosis of Influenza A Virus. Viruses. 2018; 10(11). PMC: 6266268. DOI: 10.3390/v10110650. View

14.

OKeefe B, Smee D, Turpin J, Saucedo C, Gustafson K, Mori T . Potent anti-influenza activity of cyanovirin-N and interactions with viral hemagglutinin. Antimicrob Agents Chemother. 2003; 47(8):2518-25. PMC: 166092. DOI: 10.1128/AAC.47.8.2518-2525.2003. View

15.

Arita Y, Nishimura S, Ishitsuka R, Kishimoto T, Ikenouchi J, Ishii K . Targeting cholesterol in a liquid-disordered environment by theonellamides modulates cell membrane order and cell shape. Chem Biol. 2015; 22(5):604-10. DOI: 10.1016/j.chembiol.2015.04.011. View

16.

Kusumi A, Suzuki K . Toward understanding the dynamics of membrane-raft-based molecular interactions. Biochim Biophys Acta. 2005; 1746(3):234-51. DOI: 10.1016/j.bbamcr.2005.10.001. View

17.

Srinivasan K, Raman R, Jayaraman A, Viswanathan K, Sasisekharan R . Quantitative characterization of glycan-receptor binding of H9N2 influenza A virus hemagglutinin. PLoS One. 2013; 8(4):e59550. PMC: 3634032. DOI: 10.1371/journal.pone.0059550. View

18.

Molinari N, Ortega-Sanchez I, Messonnier M, Thompson W, Wortley P, Weintraub E . The annual impact of seasonal influenza in the US: measuring disease burden and costs. Vaccine. 2007; 25(27):5086-96. DOI: 10.1016/j.vaccine.2007.03.046. View

19.

McKimm-Breschkin J . Resistance of influenza viruses to neuraminidase inhibitors--a review. Antiviral Res. 2000; 47(1):1-17. DOI: 10.1016/s0166-3542(00)00103-0. View

20.

Sarangi N, Prabhakaran A, Keyes T . Multimodal Investigation into the Interaction of Quinacrine with Microcavity-Supported Lipid Bilayers. Langmuir. 2022; 38(20):6411-6424. PMC: 9134496. DOI: 10.1021/acs.langmuir.2c00524. View