TY - JOUR
T1 - A simple goniometer-compatible flow cell for serial synchrotron X-ray crystallography
AU - Ghosh, Swagatha
AU - Zorić, Doris
AU - Dahl, Peter
AU - Bjelčić, Monika
AU - Johannesson, Jonatan
AU - Sandelin, Emil
AU - Borjesson, Per
AU - Björling, Alexander
AU - Banacore, Analia
AU - Edlund, Petra
AU - Aurelius, Oskar
AU - Milas, Mirko
AU - Nan, Jie
AU - Shilova, Anastasya
AU - Gonzalez, Ana
AU - Mueller, Uwe
AU - Brändén, Gisela
AU - Neutze, Richard
PY - 2023/3/9
Y1 - 2023/3/9
N2 - Serial femtosecond crystallography was initially developed for room-temperature X-ray diffraction studies of macromolecules at X-ray free electron lasers. When combined with tools that initiate biological reactions within microcrystals, time-resolved serial crystallography allows the study of structural changes that occur during an enzyme catalytic reaction. Serial synchrotron X-ray crystallography (SSX), which extends serial crystallography methods to synchrotron radiation sources, is expanding the scientific community using serial diffraction methods. This report presents a simple flow cell that can be used to deliver microcrystals across an X-ray beam during SSX studies. This device consists of an X-ray transparent glass capillary mounted on a goniometer-compatible 3D-printed support and is connected to a syringe pump via lightweight tubing. This flow cell is easily mounted and aligned, and it is disposable so can be rapidly replaced when blocked. This system was demonstrated by collecting SSX data at MAX IV Laboratory from microcrystals of the integral membrane protein cytochrome c oxidase from Thermus thermophilus, from which an X-ray structure was determined to 2.12 Å resolution. This simple SSX platform may help to lower entry barriers for non-expert users of SSX.
AB - Serial femtosecond crystallography was initially developed for room-temperature X-ray diffraction studies of macromolecules at X-ray free electron lasers. When combined with tools that initiate biological reactions within microcrystals, time-resolved serial crystallography allows the study of structural changes that occur during an enzyme catalytic reaction. Serial synchrotron X-ray crystallography (SSX), which extends serial crystallography methods to synchrotron radiation sources, is expanding the scientific community using serial diffraction methods. This report presents a simple flow cell that can be used to deliver microcrystals across an X-ray beam during SSX studies. This device consists of an X-ray transparent glass capillary mounted on a goniometer-compatible 3D-printed support and is connected to a syringe pump via lightweight tubing. This flow cell is easily mounted and aligned, and it is disposable so can be rapidly replaced when blocked. This system was demonstrated by collecting SSX data at MAX IV Laboratory from microcrystals of the integral membrane protein cytochrome c oxidase from Thermus thermophilus, from which an X-ray structure was determined to 2.12 Å resolution. This simple SSX platform may help to lower entry barriers for non-expert users of SSX.
KW - cytochrome c oxidase
KW - goniometer-compatible flow cells
KW - macromolecular crystallography
KW - serial synchrotron X-ray crystallography
U2 - 10.1107/S1600576723001036
DO - 10.1107/S1600576723001036
M3 - Article
C2 - 37032973
AN - SCOPUS:85159150434
SN - 0021-8898
VL - 56
SP - 449
EP - 460
JO - Journal of Applied Crystallography
JF - Journal of Applied Crystallography
IS - Pt 2
ER -