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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...

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Autores principales: Barclay, Abigail, Tidemand Johansen, Nicolai, Tidemand, Frederik Grønbæk, Arleth, Lise, Pedersen, Martin Cramer
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2022
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.
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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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