Rapid screening of in cellulo grown protein crystals via a small-angle X-ray scattering/X-ray powder diffraction synergistic approach

Lahey-Rudolph J, Schönherr R, Jeffries C, Blanchet C, Boger J, Ferreira Ramos A, Riekehr W, Triandafillidis D, Valmas A, Margiolaki I, Svergun D, Redecke L, Journal of Applied Crystallography 53(5) (2020) DOI

SASDHZ5 – In cellulo inosine-5'-monophosphate dehydrogenase (IMPDH) protein crystals recombinantly expressed within High Five insect cells

Inosine-5'-monophosphate dehydrogenase
MWexperimental 59 kDa
MWexpected 59 kDa
log I(s) 3.50×105 3.50×104 3.50×103 3.50×102
Inosine-5'-monophosphate dehydrogenase small angle scattering data  s, nm-1
Inosine-5'-monophosphate dehydrogenase Kratky plot 1.104 0 3 sRg

Data validation

There are no models related to this curve.

Synchrotron SAXS data from High Five cell cultures containing in cellulo IMPDH protein crystals were collected on the EMBL P12 beam line at PETRA III (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Data were collected at 7°C using a fixed sample position (no sample flow) in a 1.8 mm capillary. 40 successive 0.045 second frames were collected; the individual 2D images were summed (1.8 s total exposure) and then normalized to the intensity of the transmitted beam and radially averaged. The scattering of the solvent-blank was subtracted.

The High Five cell cultures were suspended in TBS (20 mM Tris, 150 mM NaCl), pH 7, that was used for solvent-background scattering subtraction. Data validation metrics do not apply for this entry (Rg, I(0), MW, etc). The quoted 'experimental MW' is that of the monomeric protein calculated from the amino acid sequence.

Inosine-5'-monophosphate dehydrogenase (IMPDH)
Mol. type   Protein
Organism   Trypanosoma brucei brucei
Olig. state   Unknown
Mon. MW   59.1 kDa
UniProt   P50098 (1-512)
Sequence   FASTA