TY  - JOUR
A1  - Schulz, Eike C.
A1  - Mehrabi, Pedram
A1  - Müller-Werkmeister, Henrike
A1  - Tellkamp, Friedjof
A1  - Jha, Ajay
A1  - Stuart, William
A1  - Persch, Elke
A1  - De Gasparo, Raoul
A1  - Diederich, François
A1  - Pai, Emil F.
A1  - Miller, R. J. Dwayne
T1  - The hit-and-return system enables efficient time-resolved serial synchrotron crystallography
JF  - Nature methods : techniques for life scientists and chemists
N2  - We present a ‘hit-and-return’ (HARE) method for time-resolved serial synchrotron crystallography with time resolution from milliseconds to seconds or longer. Timing delays are set mechanically, using the regular pattern in fixed-target crystallography chips and a translation stage system. Optical pump-probe experiments to capture intermediate structures of fluoroacetate dehalogenase binding to its ligand demonstrated that data can be collected at short (30 ms), medium (752 ms) and long (2,052 ms) intervals.
KW  - Biophysical chemistry
KW  - Enzymes
KW  - Molecular biophysics
KW  - X-ray crystallography
Y1  - 2018
U6  - https://doi.org/10.1038/s41592-018-0180-2
SN  - 1548-7091
SN  - 1548-7105
VL  - 15
IS  - 11
SP  - 901
EP  - 904
PB  - Nature Publishing Group (London)
CY  - London
ER  - 
TY  - JOUR
A1  - Otrelo-Cardoso, Ana Rita
A1  - da Silva Correia, Marcia Alexandra
A1  - Schwuchow, Viola
A1  - Svergun, Dmitri I.
A1  - Romao, Maria Joao
A1  - Leimkühler, Silke
A1  - Santos-Silva, Teresa
T1  - Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis
JF  - International journal of molecular sciences
N2  - The periplasmic aldehyde oxidoreductase PaoABC from Escherichia coli is a molybdenum enzyme involved in detoxification of aldehydes in the cell. It is an example of an heterotrimeric enzyme of the xanthine oxidase family of enzymes which does not dimerize via its molybdenum cofactor binding domain. In order to structurally characterize PaoABC, X-ray crystallography and small angle X-ray scattering (SAXS) have been carried out. The protein crystallizes in the presence of 20% (w/v) polyethylene glycol 3350 using the hanging-drop vapour diffusion method. Although crystals were initially twinned, several experiments were done to overcome twinning and lowering the crystallization temperature (293 K to 277 K) was the solution to the problem. The non-twinned crystals used to solve the structure diffract X-rays to beyond 1.80 angstrom and belong to the C2 space group, with cell parameters a = 109.42 angstrom, b = 78.08 angstrom, c = 151.77 angstrom, = 99.77 degrees, and one molecule in the asymmetric unit. A molecular replacement solution was found for each subunit separately, using several proteins as search models. SAXS data of PaoABC were also collected showing that, in solution, the protein is also an heterotrimer.
KW  - periplasmic aldehyde oxidoreductase
KW  - X-ray crystallography
KW  - small angle X-ray scattering
KW  - crystal twinning
Y1  - 2014
U6  - https://doi.org/10.3390/ijms15022223
SN  - 1422-0067
VL  - 15
IS  - 2
SP  - 2223
EP  - 2236
PB  - MDPI
CY  - Basel
ER  -