An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2

Human angiotensin converting enzyme 2 (hACE2) mediates the cell entry of both SARS-CoV and SARS-CoV2 and can be used as a drug target. The DNA encoding the truncated hACE2 (30-356aa) was cloned into pET-28a (+) and expressed in Escherichia coli Rosetta (DE3). The recombinant hACE2 (rhACE2) was purif...

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Autores principales: Gao, Xiangzheng, Liang, Keying, Mei, Shengsheng, Peng, Shanshan, Vong, Eu Gene, Zhan, Jinbiao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Inc. 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036248/
https://www.ncbi.nlm.nih.gov/pubmed/33852951
http://dx.doi.org/10.1016/j.pep.2021.105889
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author Gao, Xiangzheng
Liang, Keying
Mei, Shengsheng
Peng, Shanshan
Vong, Eu Gene
Zhan, Jinbiao
author_facet Gao, Xiangzheng
Liang, Keying
Mei, Shengsheng
Peng, Shanshan
Vong, Eu Gene
Zhan, Jinbiao
author_sort Gao, Xiangzheng
collection PubMed
description Human angiotensin converting enzyme 2 (hACE2) mediates the cell entry of both SARS-CoV and SARS-CoV2 and can be used as a drug target. The DNA encoding the truncated hACE2 (30-356aa) was cloned into pET-28a (+) and expressed in Escherichia coli Rosetta (DE3). The recombinant hACE2 (rhACE2) was purified by affinity chromatography on a Ni-NTA column and characterized with SDS-PAGE and Western blot. The binding activity of rhACE2 to Spike protein of SARS-CoV2 was evaluated in S protein-overexpressed HEK293A cells (HEK293A-SP cells) through flow cytometry. The prokaryotic expression system is able to produce approximately 75 mg protein per liter, which would be useful for infection mechanism study, and drug screening and development of SARS-CoV2.
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spelling pubmed-80362482021-04-12 An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2 Gao, Xiangzheng Liang, Keying Mei, Shengsheng Peng, Shanshan Vong, Eu Gene Zhan, Jinbiao Protein Expr Purif Article Human angiotensin converting enzyme 2 (hACE2) mediates the cell entry of both SARS-CoV and SARS-CoV2 and can be used as a drug target. The DNA encoding the truncated hACE2 (30-356aa) was cloned into pET-28a (+) and expressed in Escherichia coli Rosetta (DE3). The recombinant hACE2 (rhACE2) was purified by affinity chromatography on a Ni-NTA column and characterized with SDS-PAGE and Western blot. The binding activity of rhACE2 to Spike protein of SARS-CoV2 was evaluated in S protein-overexpressed HEK293A cells (HEK293A-SP cells) through flow cytometry. The prokaryotic expression system is able to produce approximately 75 mg protein per liter, which would be useful for infection mechanism study, and drug screening and development of SARS-CoV2. Elsevier Inc. 2021-08 2021-04-11 /pmc/articles/PMC8036248/ /pubmed/33852951 http://dx.doi.org/10.1016/j.pep.2021.105889 Text en © 2021 Elsevier Inc. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Gao, Xiangzheng
Liang, Keying
Mei, Shengsheng
Peng, Shanshan
Vong, Eu Gene
Zhan, Jinbiao
An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2
title An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2
title_full An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2
title_fullStr An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2
title_full_unstemmed An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2
title_short An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2
title_sort efficient system to generate truncated human angiotensin converting enzyme 2 (hace2) capable of binding rbd and spike protein of sars-cov2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036248/
https://www.ncbi.nlm.nih.gov/pubmed/33852951
http://dx.doi.org/10.1016/j.pep.2021.105889
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