Physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the SD1 domain produced in rice against HIV-1

Autor: Victoria Armario-Najera, Amaya Blanco-Perera, Shilpa R. Shenoy, Yi Sun, Silvia Marfil, Jordana Muñoz-Basagoiti, Daniel Perez-Zsolt, Julià Blanco, Nuria Izquierdo-Useros, Teresa Capell, Barry R. O’Keefe, Paul Christou
Rok vydání: 2022
Předmět:
Zdroj: Plant Cell Reports
r-IGTP. Repositorio Institucional de Producción Científica del Instituto de Investigación Germans Trias i Pujol
instname
Repositorio Abierto de la UdL
Universitad de Lleida
ISSN: 0721-7714
Popis: Scytovirin (SVN) is an HIV-neutralizing lectin that features two structural domains (SD1 and SD2) that bind to HIV-1 envelope glycoproteins. We expressed SD1 in rice seeds as a potential large-scale production platform and confirmed that rice-derived SD1 binds the HIV-1 envelope glycoprotein gp120 in vitro. We analyzed the thermodynamic properties of SD1 compared to full-size SVN (produced in E. coli) by isothermal titration and differential scanning calorimetry to characterize the specific interactions between SVN/SD1 and gp120 as well as to high-mannose oligosaccharides. SVN bound with moderate affinity (K-d = 1.5 mu M) to recombinant gp120, with 2.5-fold weaker affinity to nonamannoside (K-d of 3.9 mu M), and with tenfold weaker affinity to tetramannoside (13.8 mu M). The melting temperature (T-m) of full-size SVN was 59.1 degrees C and the enthalpy of unfolding (Delta H-unf) was 16.4 kcal/mol, but the T-m fell when SVN bound to nonamannoside (56.5 degrees C) and twice as much energy was required for unfolding (Delta H-unf =33.5 kcal/mol). Interestingly, binding to tetramannoside destabilized the structure of SD1 (Delta T-m similar to 11.5 degrees C) and doubled the enthalpy of unfolding, suggesting a dimerization event. The similar melting phenomenon shared by SVN and SD1 in the presence of oligomannose confirmed their conserved oligosaccharide-binding mechanisms. SD1 expressed in transgenic rice was able to neutralize HIV-1 in vitro. SD1 expressed in rice, therefore, is suitable as a microbicide component. Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work was supported by Agencia de Gestio d'Ajuts Universitaris i de Recerca (AGAUR), Departament d'Empresa i Coneixement de la Generalitat de Catalunya (PANDEMIES 2020), MINECO, Spain (AGL2017-85377-R to T. Capell), Generalitat de Catalunya Grant 2017 SGR 828 to the Agricultural Biotechnology and Bioeconomy Unit (ABBU), CERCA Programme/Generalitat de Catalunya 2017 SGR 252 to IrsiCaixa, EU Pharma-Factory grant agreement 774078 to P. Christou and J. Blanco. A. Blanco-Perera was supported by a UdL fellowship. This research was supported by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government. This project has been funded in whole or in part with federal funds from the National Cancer Institute, National Institutes of Health, under contract HHSN261201800001I.
Databáze: OpenAIRE
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