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

SASDHY5 – In cellulo luciferase protein crystals recombinantly expressed within High Five insect cells

Photinus pyralis firefly luciferase
MWexperimental 61 kDa
MWexpected 61 kDa
log I(s) 3.50×105 3.50×104 3.50×103 3.50×102
Photinus pyralis firefly luciferase small angle scattering data  s, nm-1
Photinus pyralis firefly luciferase 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 luciferase 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.

Photinus pyralis firefly luciferase (PpyLuc1)
Mol. type   Protein
Organism   Photinus pyralis
Olig. state   Unknown
Mon. MW   60.9 kDa
UniProt   H1AD96 (1-550)
Sequence   FASTA