tetano
Editor, Senior Moderator
ACS Omega
. 2023 Jul 6;8(28):25009-25019.
doi: 10.1021/acsomega.3c01625. eCollection 2023 Jul 18. In Vitro Reconstitution and Analysis of SARS-CoV-2/Host Protein-Protein Interactions
Shayli Varasteh Moradi[SUP] 1 [/SUP], Yue Wu[SUP] 1 [/SUP], Patricia Walden[SUP] 1 [/SUP], Zhenling Cui[SUP] 1 [/SUP], Wayne A Johnston[SUP] 1 [/SUP], Dmitri Petrov[SUP] 2 [/SUP], Kirill Alexandrov[SUP] 1 [/SUP]
Affiliations
The emergence of viral threats such as Ebola, ZIKA, and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) requires a rapid and efficient approach for elucidating mechanisms of pathogenesis and development of therapeutics. In this context, cell-free protein synthesis (CFPS) holds a promise to resolve the bottlenecks of multiplexed protein production and interaction analysis among host and pathogen proteins. Here, we applied a eukaryotic CFPS system based on Leishmania tarentolae extract (LTE) protein expression in combination with AlphaLISA proximity-based protein interaction technology to identify intraviral and viral-human protein interactions of SARS-CoV-2 virus that can potentially be targeted by the existing or novel antiviral therapeutics. We produced and tested 54 putative human-viral protein pairs in vitro and identified 45 direct binary protein interactions. As a casing example of the assay's suitability for drug development applications, we analyzed the effect of a putative biologic on the human angiotensin-converting enzyme 2/receptor-binding domain (hACE2/RBD) interaction. This suggests that the presented pathogen characterization platform can facilitate the development of new therapeutic agents.
. 2023 Jul 6;8(28):25009-25019.
doi: 10.1021/acsomega.3c01625. eCollection 2023 Jul 18. In Vitro Reconstitution and Analysis of SARS-CoV-2/Host Protein-Protein Interactions
Shayli Varasteh Moradi[SUP] 1 [/SUP], Yue Wu[SUP] 1 [/SUP], Patricia Walden[SUP] 1 [/SUP], Zhenling Cui[SUP] 1 [/SUP], Wayne A Johnston[SUP] 1 [/SUP], Dmitri Petrov[SUP] 2 [/SUP], Kirill Alexandrov[SUP] 1 [/SUP]
Affiliations
- PMID: 37483225
- PMCID: PMC10357528
- DOI: 10.1021/acsomega.3c01625
The emergence of viral threats such as Ebola, ZIKA, and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) requires a rapid and efficient approach for elucidating mechanisms of pathogenesis and development of therapeutics. In this context, cell-free protein synthesis (CFPS) holds a promise to resolve the bottlenecks of multiplexed protein production and interaction analysis among host and pathogen proteins. Here, we applied a eukaryotic CFPS system based on Leishmania tarentolae extract (LTE) protein expression in combination with AlphaLISA proximity-based protein interaction technology to identify intraviral and viral-human protein interactions of SARS-CoV-2 virus that can potentially be targeted by the existing or novel antiviral therapeutics. We produced and tested 54 putative human-viral protein pairs in vitro and identified 45 direct binary protein interactions. As a casing example of the assay's suitability for drug development applications, we analyzed the effect of a putative biologic on the human angiotensin-converting enzyme 2/receptor-binding domain (hACE2/RBD) interaction. This suggests that the presented pathogen characterization platform can facilitate the development of new therapeutic agents.