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

SASDFD6 – Protein kinase YopO - cytoplasmic 1 actin complex

Protein kinase YopO
Actin, cytoplasmic 1
MWexperimental 109 kDa
MWexpected 105 kDa
VPorod 147 nm3
log I(s) 6.11×102 6.11×101 6.11×100 6.11×10-1
Protein kinase YopO Actin, cytoplasmic 1 small angle scattering data  s, nm-1
ln I(s)
Protein kinase YopO Actin, cytoplasmic 1 Guinier plot ln 6.11×102 Rg: 3.6 nm 0 (3.6 nm)-2 s2
(sRg)2I(s)/I(0)
Protein kinase YopO Actin, cytoplasmic 1 Kratky plot 1.104 0 3 sRg
p(r)
Protein kinase YopO Actin, cytoplasmic 1 pair distance distribution function Rg: 3.7 nm 0 Dmax: 12.3 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Protein kinase YopO Actin, cytoplasmic 1 CUSTOM IN-HOUSE model

log I(s)
 s, nm-1
Protein kinase YopO Actin, cytoplasmic 1 CUSTOM IN-HOUSE model

log I(s)
 s, nm-1
Protein kinase YopO Actin, cytoplasmic 1 CUSTOM IN-HOUSE model

Synchrotron SAXS data from solutions of protein kinase YopO in complex with actin 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 80.00 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.

Protein kinase YopO (YopO)
Mol. type   Protein
Organism   Yersinia enterocolitica
Olig. state   Monomer
Mon. MW   63.4 kDa
 
UniProt   Q93KQ6 (89-729)
Sequence   FASTA
 
Actin, cytoplasmic 1 (Actin)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   41.7 kDa
 
UniProt   P60709 (1-375)
Sequence   FASTA