title: In situ X-ray data collection and structure phasing of protein crystals at Structural Biology Center 19-ID.
abstract: 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.
authors.affiliation: Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, USA.
authors.forename: Karolina
authors.initials: K
authors.lastname: Michalska
authors.affiliation: Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, USA.
authors.forename: Kemin
authors.initials: K
authors.lastname: Tan
authors.affiliation: Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, USA.
authors.forename: Changsoo
authors.initials: C
authors.lastname: Chang
authors.affiliation: Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: Hatzos-Skintges
authors.affiliation: Structural Biology Center, Biosciences Division, Argonne National Laboratory, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Molitsky
authors.affiliation: Structural Biology Center, Biosciences Division, Argonne National Laboratory, USA.
authors.forename: Randy
authors.initials: R
authors.lastname: Alkire
authors.affiliation: Midwest Center for Structural Genomics, Biosciences Division, Argonne National Laboratory, USA.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Joachimiak
mesh_terms: D003198:Computer Simulation; D003460:Crystallization / Q000295:instrumentation*; D018360:Crystallography, X-Ray / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008956:Models, Chemical; D008958:Models, Molecular; D044367:Phase Transition; D040081:Protein Array Analysis / Q000295:instrumentation*; D011506:Proteins / Q000737:chemistry* / Q000648:ultrastructure*
chemical_list: D011506:Proteins
keywords: crystallization plate; in situ; synchrotron data collection