Open AccessIn situ X‐ray data collection and structure phasing of protein crystals at Structural Biology Center 19‐ID
Author(s) -
Michalska Karolina,
Tan Kemin,
Chang Changsoo,
Li Hui,
HatzosSkintges Catherine,
Molitsky Michael,
Alkire Randy,
Joachimiak Andrzej
Publication year - 2015
Publication title -
journal of synchrotron radiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.172
H-Index - 99
ISSN - 1600-5775
DOI - 10.1107/s1600577515016598
Subject(s) - mosaicity , protein crystallization , data collection , beamline , phaser , in situ , advanced photon source , crystallization , computer science , crystal (programming language) , materials science , optics , diffraction , x ray crystallography , crystallography , physics , chemistry , beam (structure) , statistics , mathematics , meteorology , thermodynamics , programming language
A prototype of a 96‐well plate scanner for in situ data collection has been developed at the Structural Biology Center (SBC) beamline 19‐ID, located at the Advanced Photon Source, USA. The applicability of this instrument for protein crystal diffraction screening and data collection at ambient temperature has been demonstrated. Several different protein crystals, including selenium‐labeled, were used for data collection and successful SAD phasing. Without the common procedure of crystal handling and subsequent cryo‐cooling for data collection at T = 100 K, crystals in a crystallization buffer show remarkably low mosaicity (<0.1°) until deterioration by radiation damage occurs. Data presented here show that cryo‐cooling can cause some unexpected structural changes. Based on the results of this study, the integration of the plate scanner into the 19‐ID end‐station with automated controls is being prepared. With improvement of hardware and software, in situ data collection will become available for the SBC user program including remote access.