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Single-site glycine-specific labeling of proteins
Labeling of native proteins invites interest from diverse segments of science. However, there remains the significant unmet challenge in precise labeling at a single site of a protein. Here, we report the site-specific labeling of natural or easy-to-engineer N-terminus Gly in proteins with remarkabl...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6557831/ https://www.ncbi.nlm.nih.gov/pubmed/31182711 http://dx.doi.org/10.1038/s41467-019-10503-7 |
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author | Purushottam, Landa Adusumalli, Srinivasa Rao Singh, Usha Unnikrishnan, V. B. Rawale, Dattatraya Gautam Gujrati, Mansi Mishra, Ram Kumar Rai, Vishal |
author_facet | Purushottam, Landa Adusumalli, Srinivasa Rao Singh, Usha Unnikrishnan, V. B. Rawale, Dattatraya Gautam Gujrati, Mansi Mishra, Ram Kumar Rai, Vishal |
author_sort | Purushottam, Landa |
collection | PubMed |
description | Labeling of native proteins invites interest from diverse segments of science. However, there remains the significant unmet challenge in precise labeling at a single site of a protein. Here, we report the site-specific labeling of natural or easy-to-engineer N-terminus Gly in proteins with remarkable efficiency and selectivity. The method generates a latent nucleophile from N-terminus imine that reacts with an aldehyde to deliver an aminoalcohol under physiological conditions. It differentiates N-Gly as a unique target amongst other proteinogenic amino acids. The method allows single-site labeling of proteins in isolated form and extends to lysed cells. It administers an orthogonal aldehyde group primed for late-stage tagging with an affinity tag, (19)F NMR probe, and a fluorophore. A user-friendly protocol delivers analytically pure tagged proteins. The mild reaction conditions do not alter the structure and function of the protein. The cellular uptake of fluorophore-tagged insulin and its ability to activate the insulin-receptor mediated signaling remains unperturbed. |
format | Online Article Text |
id | pubmed-6557831 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-65578312019-06-21 Single-site glycine-specific labeling of proteins Purushottam, Landa Adusumalli, Srinivasa Rao Singh, Usha Unnikrishnan, V. B. Rawale, Dattatraya Gautam Gujrati, Mansi Mishra, Ram Kumar Rai, Vishal Nat Commun Article Labeling of native proteins invites interest from diverse segments of science. However, there remains the significant unmet challenge in precise labeling at a single site of a protein. Here, we report the site-specific labeling of natural or easy-to-engineer N-terminus Gly in proteins with remarkable efficiency and selectivity. The method generates a latent nucleophile from N-terminus imine that reacts with an aldehyde to deliver an aminoalcohol under physiological conditions. It differentiates N-Gly as a unique target amongst other proteinogenic amino acids. The method allows single-site labeling of proteins in isolated form and extends to lysed cells. It administers an orthogonal aldehyde group primed for late-stage tagging with an affinity tag, (19)F NMR probe, and a fluorophore. A user-friendly protocol delivers analytically pure tagged proteins. The mild reaction conditions do not alter the structure and function of the protein. The cellular uptake of fluorophore-tagged insulin and its ability to activate the insulin-receptor mediated signaling remains unperturbed. Nature Publishing Group UK 2019-06-10 /pmc/articles/PMC6557831/ /pubmed/31182711 http://dx.doi.org/10.1038/s41467-019-10503-7 Text en © The Author(s) 2019 Open Access 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/. |
spellingShingle | Article Purushottam, Landa Adusumalli, Srinivasa Rao Singh, Usha Unnikrishnan, V. B. Rawale, Dattatraya Gautam Gujrati, Mansi Mishra, Ram Kumar Rai, Vishal Single-site glycine-specific labeling of proteins |
title | Single-site glycine-specific labeling of proteins |
title_full | Single-site glycine-specific labeling of proteins |
title_fullStr | Single-site glycine-specific labeling of proteins |
title_full_unstemmed | Single-site glycine-specific labeling of proteins |
title_short | Single-site glycine-specific labeling of proteins |
title_sort | single-site glycine-specific labeling of proteins |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6557831/ https://www.ncbi.nlm.nih.gov/pubmed/31182711 http://dx.doi.org/10.1038/s41467-019-10503-7 |
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