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

KEY MESSAGE: Rice-produced SD1 retains its physicochemical properties and provides efficient pre-exposure HIV-1 prophylaxis against infection in vitro. ABSTRACT: Scytovirin (SVN) is an HIV-neutralizing lectin that features two structural domains (SD1 and SD2) that bind to HIV-1 envelope glycoprotein...

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Autores principales: Armario-Najera, Victoria, Blanco-Perera, Amaya, Shenoy, Shilpa R., Sun, Yi, Marfil, Silvia, Muñoz-Basagoiti, Jordana, Perez-Zsolt, Daniel, Blanco, Julià, Izquierdo-Useros, Nuria, Capell, Teresa, O’Keefe, Barry R., Christou, Paul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9034974/
https://www.ncbi.nlm.nih.gov/pubmed/35178612
http://dx.doi.org/10.1007/s00299-022-02834-5
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author Armario-Najera, Victoria
Blanco-Perera, Amaya
Shenoy, Shilpa R.
Sun, Yi
Marfil, Silvia
Muñoz-Basagoiti, Jordana
Perez-Zsolt, Daniel
Blanco, Julià
Izquierdo-Useros, Nuria
Capell, Teresa
O’Keefe, Barry R.
Christou, Paul
author_facet Armario-Najera, Victoria
Blanco-Perera, Amaya
Shenoy, Shilpa R.
Sun, Yi
Marfil, Silvia
Muñoz-Basagoiti, Jordana
Perez-Zsolt, Daniel
Blanco, Julià
Izquierdo-Useros, Nuria
Capell, Teresa
O’Keefe, Barry R.
Christou, Paul
author_sort Armario-Najera, Victoria
collection PubMed
description KEY MESSAGE: Rice-produced SD1 retains its physicochemical properties and provides efficient pre-exposure HIV-1 prophylaxis against infection in vitro. ABSTRACT: 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 µM) to recombinant gp120, with 2.5-fold weaker affinity to nonamannoside (K(d) of 3.9 µM), and with tenfold weaker affinity to tetramannoside (13.8 µM). The melting temperature (T(m)) of full-size SVN was 59.1 °C and the enthalpy of unfolding (ΔH(unf)) was 16.4 kcal/mol, but the T(m) fell when SVN bound to nonamannoside (56.5 °C) and twice as much energy was required for unfolding (ΔH(unf) = 33.5 kcal/mol). Interestingly, binding to tetramannoside destabilized the structure of SD1 (ΔT(m) ~ 11.5 °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.
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spelling pubmed-90349742022-05-06 Physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the SD1 domain produced in rice against HIV-1 Armario-Najera, Victoria Blanco-Perera, Amaya Shenoy, Shilpa R. Sun, Yi Marfil, Silvia Muñoz-Basagoiti, Jordana Perez-Zsolt, Daniel Blanco, Julià Izquierdo-Useros, Nuria Capell, Teresa O’Keefe, Barry R. Christou, Paul Plant Cell Rep Original Article KEY MESSAGE: Rice-produced SD1 retains its physicochemical properties and provides efficient pre-exposure HIV-1 prophylaxis against infection in vitro. ABSTRACT: 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 µM) to recombinant gp120, with 2.5-fold weaker affinity to nonamannoside (K(d) of 3.9 µM), and with tenfold weaker affinity to tetramannoside (13.8 µM). The melting temperature (T(m)) of full-size SVN was 59.1 °C and the enthalpy of unfolding (ΔH(unf)) was 16.4 kcal/mol, but the T(m) fell when SVN bound to nonamannoside (56.5 °C) and twice as much energy was required for unfolding (ΔH(unf) = 33.5 kcal/mol). Interestingly, binding to tetramannoside destabilized the structure of SD1 (ΔT(m) ~ 11.5 °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. Springer Berlin Heidelberg 2022-02-18 2022 /pmc/articles/PMC9034974/ /pubmed/35178612 http://dx.doi.org/10.1007/s00299-022-02834-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Original Article
Armario-Najera, Victoria
Blanco-Perera, Amaya
Shenoy, Shilpa R.
Sun, Yi
Marfil, Silvia
Muñoz-Basagoiti, Jordana
Perez-Zsolt, Daniel
Blanco, Julià
Izquierdo-Useros, Nuria
Capell, Teresa
O’Keefe, Barry R.
Christou, Paul
Physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the SD1 domain produced in rice against HIV-1
title Physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the SD1 domain produced in rice against HIV-1
title_full Physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the SD1 domain produced in rice against HIV-1
title_fullStr Physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the SD1 domain produced in rice against HIV-1
title_full_unstemmed Physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the SD1 domain produced in rice against HIV-1
title_short Physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the SD1 domain produced in rice against HIV-1
title_sort physicochemical characterization of the recombinant lectin scytovirin and microbicidal activity of the sd1 domain produced in rice against hiv-1
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9034974/
https://www.ncbi.nlm.nih.gov/pubmed/35178612
http://dx.doi.org/10.1007/s00299-022-02834-5
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