Perturbing the folding energy landscape of the bacterial immunity protein Im7 by site-specific N-linked glycosylation
N-linked glycosylation modulates protein folding and stability through a variety of mechanisms. As such there is considerable interest in the development of general rules to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in the...
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| Language: | en_US |
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National Academy of Sciences
2011
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| Online Access: | http://hdl.handle.net/1721.1/64814 https://orcid.org/0000-0002-3415-242X https://orcid.org/0000-0002-5749-7869 |
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| author | Chen, Mark Bartlett, Alice I. Nerenberg, Paul S. Friel, Claire T. Hackenberger, Christian P. R. Radford, Sheena E. Imperiali, Barbara Stultz, Collin M |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Chen, Mark Bartlett, Alice I. Nerenberg, Paul S. Friel, Claire T. Hackenberger, Christian P. R. Radford, Sheena E. Imperiali, Barbara Stultz, Collin M |
| author_sort | Chen, Mark |
| collection | MIT |
| description | N-linked glycosylation modulates protein folding and stability through a variety of mechanisms. As such there is considerable interest in the development of general rules to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in the design and development of modified proteins with advantageous properties. In this study, expressed protein ligation is used to create site-specifically glycosylated variants of the bacterial immunity protein Im7 modified with the chitobiose disaccharide (GlcNAc-GlcNAc). Glycans were introduced at seven solvent exposed sites within the Im7 sequence and the kinetic and thermodynamic consequences of N-linked glycosylation analyzed. The ΔΔG° [delta delta G superscript 0 or degree symbol] values for glycan incorporation were found to range from +5.2 to -3.8 kJ·mol-1. In several cases, glycosylation influences folding by modulating the local conformational preferences of the glycosylated sequence. These locally mediated effects are most prominent in the center of α-helices where glycosylation negatively effects folding and in compact turn motifs between segments of ordered secondary structure where glycosylation promotes folding and enhances the overall stability of the native protein. The studies also provide insight into why glycosylation is commonly identified at the transition between different types of secondary structure and when glycosylation may be used to elaborate protein structure to protect disordered sequences from proteolysis or immune system recognition. |
| first_indexed | 2024-09-23T09:59:43Z |
| format | Article |
| id | mit-1721.1/64814 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:59:43Z |
| publishDate | 2011 |
| publisher | National Academy of Sciences |
| record_format | dspace |
| spelling | mit-1721.1/648142022-09-26T15:02:23Z Perturbing the folding energy landscape of the bacterial immunity protein Im7 by site-specific N-linked glycosylation Chen, Mark Bartlett, Alice I. Nerenberg, Paul S. Friel, Claire T. Hackenberger, Christian P. R. Radford, Sheena E. Imperiali, Barbara Stultz, Collin M Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Massachusetts Institute of Technology. Department of Physics Imperiali, Barbara Chen, Mark Nerenberg, Paul S. Hackenberger, Christian P. R. Stultz, Collin M. Imperiali, Barbara N-linked glycosylation modulates protein folding and stability through a variety of mechanisms. As such there is considerable interest in the development of general rules to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in the design and development of modified proteins with advantageous properties. In this study, expressed protein ligation is used to create site-specifically glycosylated variants of the bacterial immunity protein Im7 modified with the chitobiose disaccharide (GlcNAc-GlcNAc). Glycans were introduced at seven solvent exposed sites within the Im7 sequence and the kinetic and thermodynamic consequences of N-linked glycosylation analyzed. The ΔΔG° [delta delta G superscript 0 or degree symbol] values for glycan incorporation were found to range from +5.2 to -3.8 kJ·mol-1. In several cases, glycosylation influences folding by modulating the local conformational preferences of the glycosylated sequence. These locally mediated effects are most prominent in the center of α-helices where glycosylation negatively effects folding and in compact turn motifs between segments of ordered secondary structure where glycosylation promotes folding and enhances the overall stability of the native protein. The studies also provide insight into why glycosylation is commonly identified at the transition between different types of secondary structure and when glycosylation may be used to elaborate protein structure to protect disordered sequences from proteolysis or immune system recognition. National Institutes of Health (U.S.) (GM039334) National Science Foundation (U.S.) (0821391) Biotechnology and Biological Sciences Research Council (Great Britain) (Grant BB/526502/1) Biotechnology and Biological Sciences Research Council (Great Britain) (Grant 24/B17145) 2011-07-14T18:35:22Z 2011-07-14T18:35:22Z 2010-12 2010-09 Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/64814 Chen, M. M. et al. “Perturbing the Folding Energy Landscape of the Bacterial Immunity Protein Im7 by Site-specific N-linked Glycosylation.” Proceedings of the National Academy of Sciences 107.52 (2010) : 22528-22533. https://orcid.org/0000-0002-3415-242X https://orcid.org/0000-0002-5749-7869 en_US http://dx.doi.org/10.1073/pnas.1015356107 Proceedings of the National Academy of Sciences of the United States of America Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences PNAS |
| spellingShingle | Chen, Mark Bartlett, Alice I. Nerenberg, Paul S. Friel, Claire T. Hackenberger, Christian P. R. Radford, Sheena E. Imperiali, Barbara Stultz, Collin M Perturbing the folding energy landscape of the bacterial immunity protein Im7 by site-specific N-linked glycosylation |
| title | Perturbing the folding energy landscape of the bacterial immunity protein Im7 by site-specific N-linked glycosylation |
| title_full | Perturbing the folding energy landscape of the bacterial immunity protein Im7 by site-specific N-linked glycosylation |
| title_fullStr | Perturbing the folding energy landscape of the bacterial immunity protein Im7 by site-specific N-linked glycosylation |
| title_full_unstemmed | Perturbing the folding energy landscape of the bacterial immunity protein Im7 by site-specific N-linked glycosylation |
| title_short | Perturbing the folding energy landscape of the bacterial immunity protein Im7 by site-specific N-linked glycosylation |
| title_sort | perturbing the folding energy landscape of the bacterial immunity protein im7 by site specific n linked glycosylation |
| url | http://hdl.handle.net/1721.1/64814 https://orcid.org/0000-0002-3415-242X https://orcid.org/0000-0002-5749-7869 |
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