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Dispersion from C(α) or N(H): 4D experiments for backbone resonance assignment of intrinsically disordered proteins
Resonance assignment of intrinsically disordered proteins is remarkably challenging due to scant chemical shift dispersion arising from conformational heterogeneity. The challenge is even greater if repeating segments are present in the amino acid sequence. To forward unambiguous resonance assignmen...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7080685/ https://www.ncbi.nlm.nih.gov/pubmed/31932991 http://dx.doi.org/10.1007/s10858-020-00299-w |
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author | Tossavainen, Helena Salovaara, Santeri Hellman, Maarit Ihalin, Riikka Permi, Perttu |
author_facet | Tossavainen, Helena Salovaara, Santeri Hellman, Maarit Ihalin, Riikka Permi, Perttu |
author_sort | Tossavainen, Helena |
collection | PubMed |
description | Resonance assignment of intrinsically disordered proteins is remarkably challenging due to scant chemical shift dispersion arising from conformational heterogeneity. The challenge is even greater if repeating segments are present in the amino acid sequence. To forward unambiguous resonance assignment of intrinsically disordered proteins, we present iHACANCO, HACACON and (HACA)CONCAHA, three H(α)-detected 4D experiments with C(α) as an additional dimension. In addition, we present (HACA)CON(CA)NH and (HACA)N(CA)CONH, new 4D H(α)-start, H(N)-detect experiments which have two N(H) dimensions to enhance peak dispersion in a sequential walk through C′, N(H) and H(N), and provide more accurate N(H)/H(N) chemical shifts than those that can be obtained from a crowded (1)H, (15)N-HSQC spectrum. Application of these 4D experiments is demonstrated using BilRI (165 aa), an outer-membrane intrinsically disordered protein from the opportunistic oral pathogen Aggregatibacter actinomycetemcomitans. BilRI amino acid sequence encompasses three very similar repeats with a 13-residue identical stretch in two of them. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10858-020-00299-w) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-7080685 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-70806852020-03-23 Dispersion from C(α) or N(H): 4D experiments for backbone resonance assignment of intrinsically disordered proteins Tossavainen, Helena Salovaara, Santeri Hellman, Maarit Ihalin, Riikka Permi, Perttu J Biomol NMR Article Resonance assignment of intrinsically disordered proteins is remarkably challenging due to scant chemical shift dispersion arising from conformational heterogeneity. The challenge is even greater if repeating segments are present in the amino acid sequence. To forward unambiguous resonance assignment of intrinsically disordered proteins, we present iHACANCO, HACACON and (HACA)CONCAHA, three H(α)-detected 4D experiments with C(α) as an additional dimension. In addition, we present (HACA)CON(CA)NH and (HACA)N(CA)CONH, new 4D H(α)-start, H(N)-detect experiments which have two N(H) dimensions to enhance peak dispersion in a sequential walk through C′, N(H) and H(N), and provide more accurate N(H)/H(N) chemical shifts than those that can be obtained from a crowded (1)H, (15)N-HSQC spectrum. Application of these 4D experiments is demonstrated using BilRI (165 aa), an outer-membrane intrinsically disordered protein from the opportunistic oral pathogen Aggregatibacter actinomycetemcomitans. BilRI amino acid sequence encompasses three very similar repeats with a 13-residue identical stretch in two of them. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10858-020-00299-w) contains supplementary material, which is available to authorized users. Springer Netherlands 2020-01-13 2020 /pmc/articles/PMC7080685/ /pubmed/31932991 http://dx.doi.org/10.1007/s10858-020-00299-w Text en © The Author(s) 2020 Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Tossavainen, Helena Salovaara, Santeri Hellman, Maarit Ihalin, Riikka Permi, Perttu Dispersion from C(α) or N(H): 4D experiments for backbone resonance assignment of intrinsically disordered proteins |
title | Dispersion from C(α) or N(H): 4D experiments for backbone resonance assignment of intrinsically disordered proteins |
title_full | Dispersion from C(α) or N(H): 4D experiments for backbone resonance assignment of intrinsically disordered proteins |
title_fullStr | Dispersion from C(α) or N(H): 4D experiments for backbone resonance assignment of intrinsically disordered proteins |
title_full_unstemmed | Dispersion from C(α) or N(H): 4D experiments for backbone resonance assignment of intrinsically disordered proteins |
title_short | Dispersion from C(α) or N(H): 4D experiments for backbone resonance assignment of intrinsically disordered proteins |
title_sort | dispersion from c(α) or n(h): 4d experiments for backbone resonance assignment of intrinsically disordered proteins |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7080685/ https://www.ncbi.nlm.nih.gov/pubmed/31932991 http://dx.doi.org/10.1007/s10858-020-00299-w |
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