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Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II

Advances in synchrotron technology are changing the landscape of macromolecular crystallography. The two recently opened beamlines at NSLS-II—AMX and FMX—deliver high-flux microfocus beams that open new possibilities for crystallographic data collection. They are equipped with state-of-the-art exper...

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Autores principales: Miller, Michelle S., Maheshwari, Sweta, Shi, Wuxian, Gao, Yuan, Chu, Nam, Soares, Alexei S., Cole, Philip A., Amzel, L. Mario, Fuchs, Martin R., Jakoncic, Jean, Gabelli, Sandra B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6384729/
https://www.ncbi.nlm.nih.gov/pubmed/30704096
http://dx.doi.org/10.3390/molecules24030496
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author Miller, Michelle S.
Maheshwari, Sweta
Shi, Wuxian
Gao, Yuan
Chu, Nam
Soares, Alexei S.
Cole, Philip A.
Amzel, L. Mario
Fuchs, Martin R.
Jakoncic, Jean
Gabelli, Sandra B.
author_facet Miller, Michelle S.
Maheshwari, Sweta
Shi, Wuxian
Gao, Yuan
Chu, Nam
Soares, Alexei S.
Cole, Philip A.
Amzel, L. Mario
Fuchs, Martin R.
Jakoncic, Jean
Gabelli, Sandra B.
author_sort Miller, Michelle S.
collection PubMed
description Advances in synchrotron technology are changing the landscape of macromolecular crystallography. The two recently opened beamlines at NSLS-II—AMX and FMX—deliver high-flux microfocus beams that open new possibilities for crystallographic data collection. They are equipped with state-of-the-art experimental stations and automation to allow data collection on previously intractable crystals. Optimized data collection strategies allow users to tailor crystal positioning to optimally distribute the X-ray dose over its volume. Vector data collection allows the user to define a linear trajectory along a well diffracting volume of the crystal and perform rotational data collection while moving along the vector. This is particularly well suited to long, thin crystals. We describe vector data collection of three proteins—Akt1, PI3Kα, and CDP-Chase—to demonstrate its application and utility. For smaller crystals, we describe two methods for multicrystal data collection in a single loop, either manually selecting multiple centers (using H108A-PHM as an example), or “raster-collect”, a more automated approach for a larger number of crystals (using CDP-Chase as an example).
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spelling pubmed-63847292019-02-23 Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II Miller, Michelle S. Maheshwari, Sweta Shi, Wuxian Gao, Yuan Chu, Nam Soares, Alexei S. Cole, Philip A. Amzel, L. Mario Fuchs, Martin R. Jakoncic, Jean Gabelli, Sandra B. Molecules Article Advances in synchrotron technology are changing the landscape of macromolecular crystallography. The two recently opened beamlines at NSLS-II—AMX and FMX—deliver high-flux microfocus beams that open new possibilities for crystallographic data collection. They are equipped with state-of-the-art experimental stations and automation to allow data collection on previously intractable crystals. Optimized data collection strategies allow users to tailor crystal positioning to optimally distribute the X-ray dose over its volume. Vector data collection allows the user to define a linear trajectory along a well diffracting volume of the crystal and perform rotational data collection while moving along the vector. This is particularly well suited to long, thin crystals. We describe vector data collection of three proteins—Akt1, PI3Kα, and CDP-Chase—to demonstrate its application and utility. For smaller crystals, we describe two methods for multicrystal data collection in a single loop, either manually selecting multiple centers (using H108A-PHM as an example), or “raster-collect”, a more automated approach for a larger number of crystals (using CDP-Chase as an example). MDPI 2019-01-30 /pmc/articles/PMC6384729/ /pubmed/30704096 http://dx.doi.org/10.3390/molecules24030496 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Miller, Michelle S.
Maheshwari, Sweta
Shi, Wuxian
Gao, Yuan
Chu, Nam
Soares, Alexei S.
Cole, Philip A.
Amzel, L. Mario
Fuchs, Martin R.
Jakoncic, Jean
Gabelli, Sandra B.
Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II
title Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II
title_full Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II
title_fullStr Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II
title_full_unstemmed Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II
title_short Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II
title_sort getting the most out of your crystals: data collection at the new high-flux, microfocus mx beamlines at nsls-ii
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6384729/
https://www.ncbi.nlm.nih.gov/pubmed/30704096
http://dx.doi.org/10.3390/molecules24030496
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