skip to main content

DOE PAGESDOE PAGES

Title: 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 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).
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [5] ;  [4] ;  [5] ;  [2] ;  [4] ;  [4] ;  [6]
  1. Johns Hopkins Univ., Baltimore, MD (United States). School of Medicine. Dept. of Oncology
  2. Johns Hopkins Univ., Baltimore, MD (United States). School of Medicine. Dept. of Biophysics and Biophysical Chemistry
  3. Brookhaven National Lab. (BNL), Upton, NY (United States). Energy & Photon Sciences Directorate; Case Western Reserve Univ., Cleveland, OH (United States). Case Center for Synchrotron Biosciences
  4. Brookhaven National Lab. (BNL), Upton, NY (United States). Energy & Photon Sciences Directorate
  5. Harvard Medical School and Brigham and Women’s Hospital, Boston, MA (United States). Division of Genetics. Dept. of Medicine. Dept. of Biological Chemistry and Molecular Pharmacology
  6. Johns Hopkins Univ., Baltimore, MD (United States). School of Medicine. Dept. of Oncology. Dept. of Biophysics and Biophysical Chemistry. Dept. of Medicine
Publication Date:
Report Number(s):
BNL-211430-2019-JAAM
Journal ID: ISSN 1420-3049
Grant/Contract Number:
SC0012704; BC151831; CA062924; CA74305; P41GM111244; MCB-1517522
Type:
Accepted Manuscript
Journal Name:
Molecules
Additional Journal Information:
Journal Volume: 24; Journal Issue: 3; Journal ID: ISSN 1420-3049
Publisher:
MDPI
Research Org:
Brookhaven National Lab. (BNL), Upton, NY (United States); Johns Hopkins Univ., Baltimore, MD (United States); Harvard Medical School and Brigham and Women’s Hospital, Boston, MA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); USDOD; National Inst. of Health (NIH) (United States); National Science Foundation (NSF)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; multicrystal; vector data collection; NSLS-II; PI3Kα, Akt1; CDP-Chase; PHM; microfocus; microdiffraction; nonhomogeneous crystals
OSTI Identifier:
1502802

Miller, Michelle, Maheshwari, Sweta, Shi, Wuxian, Gao, Yuan, Chu, Nam, Soares, Alexei, Cole, Philip, Amzel, L., Fuchs, Martin, Jakoncic, Jean, and Gabelli, Sandra. Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II. United States: N. p., Web. doi:10.3390/molecules24030496.
Miller, Michelle, Maheshwari, Sweta, Shi, Wuxian, Gao, Yuan, Chu, Nam, Soares, Alexei, Cole, Philip, Amzel, L., Fuchs, Martin, Jakoncic, Jean, & Gabelli, Sandra. Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II. United States. doi:10.3390/molecules24030496.
Miller, Michelle, Maheshwari, Sweta, Shi, Wuxian, Gao, Yuan, Chu, Nam, Soares, Alexei, Cole, Philip, Amzel, L., Fuchs, Martin, Jakoncic, Jean, and Gabelli, Sandra. 2019. "Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II". United States. doi:10.3390/molecules24030496. https://www.osti.gov/servlets/purl/1502802.
@article{osti_1502802,
title = {Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II},
author = {Miller, Michelle and Maheshwari, Sweta and Shi, Wuxian and Gao, Yuan and Chu, Nam and Soares, Alexei and Cole, Philip and Amzel, L. and Fuchs, Martin and Jakoncic, Jean and Gabelli, Sandra},
abstractNote = {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).},
doi = {10.3390/molecules24030496},
journal = {Molecules},
number = 3,
volume = 24,
place = {United States},
year = {2019},
month = {1}
}

Works referenced in this record:

Integration, scaling, space-group assignment and post-refinement
journal, January 2010
  • Kabsch, Wolfgang
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 133-144
  • DOI: 10.1107/S0907444909047374

Coot model-building tools for molecular graphics
journal, November 2004
  • Emsley, Paul; Cowtan, Kevin
  • Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132
  • DOI: 10.1107/S0907444904019158