Getting the Most Out of Your Crystals: Data Collection at the New High-Flux, Microfocus MX Beamlines at NSLS-II
Michelle S. Miller,
Sweta Maheshwari,
Wuxian Shi,
Yuan Gao,
Nam Chu,
Alexei S. Soares,
Philip A. Cole,
L. Mario Amzel,
Martin R. Fuchs,
Jean Jakoncic,
Sandra B. Gabelli
Affiliations
Michelle S. Miller
Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
Sweta Maheshwari
Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
Wuxian Shi
Energy & Photon Sciences Directorate, Brookhaven National Laboratory, Upton, NY 11973, USA
Yuan Gao
Energy & Photon Sciences Directorate, Brookhaven National Laboratory, Upton, NY 11973, USA
Nam Chu
Division of Genetics, Departments of Medicine and Biological Chemistry and Molecular Pharmacology, Harvard Medical School and Brigham and Women’s Hospital, Boston, MA 02115, USA
Alexei S. Soares
Energy & Photon Sciences Directorate, Brookhaven National Laboratory, Upton, NY 11973, USA
Philip A. Cole
Division of Genetics, Departments of Medicine and Biological Chemistry and Molecular Pharmacology, Harvard Medical School and Brigham and Women’s Hospital, Boston, MA 02115, USA
L. Mario Amzel
Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
Martin R. Fuchs
Energy & Photon Sciences Directorate, Brookhaven National Laboratory, Upton, NY 11973, USA
Jean Jakoncic
Energy & Photon Sciences Directorate, Brookhaven National Laboratory, Upton, NY 11973, USA
Sandra B. Gabelli
Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
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).
