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Global fitting of multiple data frames from SEC–SAXS to investigate the structure of next-generation nanodiscs
The combination of online size-exclusion chromatography and small-angle X-ray scattering (SEC–SAXS) is rapidly becoming a key technique for structural investigations of elaborate biophysical samples in solution. Here, a novel model-refinement strategy centred around the technique is outlined and its...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8972807/ https://www.ncbi.nlm.nih.gov/pubmed/35362471 http://dx.doi.org/10.1107/S2059798322001838 |
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author | Barclay, Abigail Tidemand Johansen, Nicolai Tidemand, Frederik Grønbæk Arleth, Lise Pedersen, Martin Cramer |
author_facet | Barclay, Abigail Tidemand Johansen, Nicolai Tidemand, Frederik Grønbæk Arleth, Lise Pedersen, Martin Cramer |
author_sort | Barclay, Abigail |
collection | PubMed |
description | The combination of online size-exclusion chromatography and small-angle X-ray scattering (SEC–SAXS) is rapidly becoming a key technique for structural investigations of elaborate biophysical samples in solution. Here, a novel model-refinement strategy centred around the technique is outlined and its utility is demonstrated by analysing data series from several SEC–SAXS experiments on phospholipid bilayer nanodiscs. Using this method, a single model was globally refined against many frames from the same data series, thereby capturing the frame-to-frame tendencies of the irradiated sample. These are compared with models refined in the traditional manner, in which refinement is based on the average profile of a set of consecutive frames from the same data series without an in-depth comparison of individual frames. This is considered to be an attractive model-refinement scheme as it considerably lowers the total number of parameters refined from the data series, produces tendencies that are automatically consistent between frames, and utilizes a considerably larger portion of the recorded data than is often performed in such experiments. Additionally, a method is outlined for correcting a measured UV absorption signal by accounting for potential peak broadening by the experimental setup. |
format | Online Article Text |
id | pubmed-8972807 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-89728072022-04-28 Global fitting of multiple data frames from SEC–SAXS to investigate the structure of next-generation nanodiscs Barclay, Abigail Tidemand Johansen, Nicolai Tidemand, Frederik Grønbæk Arleth, Lise Pedersen, Martin Cramer Acta Crystallogr D Struct Biol Research Papers The combination of online size-exclusion chromatography and small-angle X-ray scattering (SEC–SAXS) is rapidly becoming a key technique for structural investigations of elaborate biophysical samples in solution. Here, a novel model-refinement strategy centred around the technique is outlined and its utility is demonstrated by analysing data series from several SEC–SAXS experiments on phospholipid bilayer nanodiscs. Using this method, a single model was globally refined against many frames from the same data series, thereby capturing the frame-to-frame tendencies of the irradiated sample. These are compared with models refined in the traditional manner, in which refinement is based on the average profile of a set of consecutive frames from the same data series without an in-depth comparison of individual frames. This is considered to be an attractive model-refinement scheme as it considerably lowers the total number of parameters refined from the data series, produces tendencies that are automatically consistent between frames, and utilizes a considerably larger portion of the recorded data than is often performed in such experiments. Additionally, a method is outlined for correcting a measured UV absorption signal by accounting for potential peak broadening by the experimental setup. International Union of Crystallography 2022-03-11 /pmc/articles/PMC8972807/ /pubmed/35362471 http://dx.doi.org/10.1107/S2059798322001838 Text en © Abigail Barclay et al. 2022 https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Barclay, Abigail Tidemand Johansen, Nicolai Tidemand, Frederik Grønbæk Arleth, Lise Pedersen, Martin Cramer Global fitting of multiple data frames from SEC–SAXS to investigate the structure of next-generation nanodiscs |
title | Global fitting of multiple data frames from SEC–SAXS to investigate the structure of next-generation nanodiscs |
title_full | Global fitting of multiple data frames from SEC–SAXS to investigate the structure of next-generation nanodiscs |
title_fullStr | Global fitting of multiple data frames from SEC–SAXS to investigate the structure of next-generation nanodiscs |
title_full_unstemmed | Global fitting of multiple data frames from SEC–SAXS to investigate the structure of next-generation nanodiscs |
title_short | Global fitting of multiple data frames from SEC–SAXS to investigate the structure of next-generation nanodiscs |
title_sort | global fitting of multiple data frames from sec–saxs to investigate the structure of next-generation nanodiscs |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8972807/ https://www.ncbi.nlm.nih.gov/pubmed/35362471 http://dx.doi.org/10.1107/S2059798322001838 |
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