Thermostable exoshells fold and stabilize recombinant proteins
The expression and stabilization of recombinant proteins is fundamental to basic and applied biology. Here we have engineered a thermostable protein nanoparticle (tES) to improve both expression and stabilization of recombinant proteins using this technology. tES provides steric accommodation and ch...
| Main Authors: | , , , , , , |
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| Format: | Journal Article |
| Language: | English |
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2018
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| Online Access: | https://hdl.handle.net/10356/89629 http://hdl.handle.net/10220/44987 |
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| author | Deshpande, Siddharth Masurkar, Nihar D. Girish, Vallerinteavide Mavelli Desai, Malan Chakraborty, Goutam Chan, Juliana M. Drum, Chester L. |
| author2 | School of Chemical and Biomedical Engineering |
| author_facet | School of Chemical and Biomedical Engineering Deshpande, Siddharth Masurkar, Nihar D. Girish, Vallerinteavide Mavelli Desai, Malan Chakraborty, Goutam Chan, Juliana M. Drum, Chester L. |
| author_sort | Deshpande, Siddharth |
| collection | NTU |
| description | The expression and stabilization of recombinant proteins is fundamental to basic and applied biology. Here we have engineered a thermostable protein nanoparticle (tES) to improve both expression and stabilization of recombinant proteins using this technology. tES provides steric accommodation and charge complementation to green fluorescent protein (GFPuv), horseradish peroxidase (HRPc), and Renilla luciferase (rLuc), improving the yields of functional in vitro folding by ~100-fold. Encapsulated enzymes retain the ability to metabolize small-molecule substrates, presumably via four 4.5-nm pores present in the tES shell. GFPuv exhibits no spectral shifts in fluorescence compared to a nonencapsulated control. Thermolabile proteins internalized by tES are resistant to thermal, organic, chaotropic, and proteolytic denaturation and can be released from the tES assembly with mild pH titration followed by proteolysis. |
| first_indexed | 2024-10-01T06:00:17Z |
| format | Journal Article |
| id | ntu-10356/89629 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T06:00:17Z |
| publishDate | 2018 |
| record_format | dspace |
| spelling | ntu-10356/896292023-12-29T06:50:07Z Thermostable exoshells fold and stabilize recombinant proteins Deshpande, Siddharth Masurkar, Nihar D. Girish, Vallerinteavide Mavelli Desai, Malan Chakraborty, Goutam Chan, Juliana M. Drum, Chester L. School of Chemical and Biomedical Engineering Lee Kong Chian School of Medicine (LKCMedicine) Nanoparticles Protein Folding The expression and stabilization of recombinant proteins is fundamental to basic and applied biology. Here we have engineered a thermostable protein nanoparticle (tES) to improve both expression and stabilization of recombinant proteins using this technology. tES provides steric accommodation and charge complementation to green fluorescent protein (GFPuv), horseradish peroxidase (HRPc), and Renilla luciferase (rLuc), improving the yields of functional in vitro folding by ~100-fold. Encapsulated enzymes retain the ability to metabolize small-molecule substrates, presumably via four 4.5-nm pores present in the tES shell. GFPuv exhibits no spectral shifts in fluorescence compared to a nonencapsulated control. Thermolabile proteins internalized by tES are resistant to thermal, organic, chaotropic, and proteolytic denaturation and can be released from the tES assembly with mild pH titration followed by proteolysis. NMRC (Natl Medical Research Council, S’pore) Published version 2018-06-07T07:05:15Z 2019-12-06T17:29:52Z 2018-06-07T07:05:15Z 2019-12-06T17:29:52Z 2017 Journal Article Deshpande, S., Masurkar, N. D., Girish, V. M., Desai, M., Chakraborty, G., Chan, J. M., et al. (2017). Thermostable exoshells fold and stabilize recombinant proteins. Nature Communications, 8(1), 1442-. https://hdl.handle.net/10356/89629 http://hdl.handle.net/10220/44987 10.1038/s41467-017-01585-2 en Nature Communications © 2017 The Author(s). This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/ licenses/by/4.0/. 8 p. application/pdf |
| spellingShingle | Nanoparticles Protein Folding Deshpande, Siddharth Masurkar, Nihar D. Girish, Vallerinteavide Mavelli Desai, Malan Chakraborty, Goutam Chan, Juliana M. Drum, Chester L. Thermostable exoshells fold and stabilize recombinant proteins |
| title | Thermostable exoshells fold and stabilize recombinant proteins |
| title_full | Thermostable exoshells fold and stabilize recombinant proteins |
| title_fullStr | Thermostable exoshells fold and stabilize recombinant proteins |
| title_full_unstemmed | Thermostable exoshells fold and stabilize recombinant proteins |
| title_short | Thermostable exoshells fold and stabilize recombinant proteins |
| title_sort | thermostable exoshells fold and stabilize recombinant proteins |
| topic | Nanoparticles Protein Folding |
| url | https://hdl.handle.net/10356/89629 http://hdl.handle.net/10220/44987 |
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