Studying Conformational Changes of the Yersinia Type-III-Secretion Effector YopO in Solution by Integrative Structural Biology.

Peter MF, Tuukkanen AT, Heubach CA, Selsam A, Duthie FG, Svergun DI, Schiemann O, Hagelueken G, Structure 27(9):1416-1426.e3 (2019) Europe PMC

SASDFC6 – Wild type protein kinase YopO

Protein kinase YopO
MWexperimental 69 kDa
MWexpected 63 kDa
VPorod 119 nm3
log I(s) 2.67×102 2.67×101 2.67×100 2.67×10-1
Protein kinase YopO small angle scattering data  s, nm-1
ln I(s)
Protein kinase YopO Guinier plot ln 2.68×102 Rg: 3.3 nm 0 (3.3 nm)-2 s2
(sRg)2I(s)/I(0)
Protein kinase YopO Kratky plot 1.104 0 √3 sRg
p(r)
Protein kinase YopO pair distance distribution function Rg: 3.4 nm 0 Dmax: 11.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Protein kinase YopO CUSTOM IN-HOUSE model
Synchrotron SAXS data from solutions of protein kinase YopO in 10 mM Tris-HCl, 50 mM NaCl, pH 8 were collected on the EMBL P12 beam line at the PETRA III (Hamburg, Germany) using a Pilatus 2M 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). In-line size-exclusion chromatography (SEC) SAXS was employed. The SEC parameters were as follows: A 80 μl sample at 5.5 mg/ml was injected onto a GE Superdex 200 Increase 10/300 column at 20°C. 3600 successive 1 second frames were collected through the SEC elution. The data from the sample peak were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Flow rate = UNKNOWN

Protein kinase YopO (YopO)
Mol. type   Protein
Organism   Yersinia enterocolitica
Olig. state   Monomer
Mon. MW   63.4 kDa
 
UniProt   Q93KQ6 (89-729)
Sequence   FASTA