Structure of the NPr:EINNtr Complex: Mechanism for Specificity in Paralogous Phosphotransferase Systems.

Strickland M, Stanley AM, Wang G, Botos I, Schwieters CD, Buchanan SK, Peterkofsky A, Tjandra N, Structure 24(12):2127-2137 (2016) Europe PMC

SASDBZ5 – Enzyme I-Ntr (residues 170-424) in complex with NPr (residues 1-85)

Phosphocarrier protein NPr
Phosphoenolpyruvate-protein phosphotransferase PtsP
MWI(0) 22 kDa
MWexpected 38 kDa
VPorod 52 nm3
log I(s) 2.83×100 2.83×10-1 2.83×10-2 2.83×10-3
Phosphocarrier protein NPr Phosphoenolpyruvate-protein phosphotransferase PtsP small angle scattering data  s, nm-1
ln I(s)
Phosphocarrier protein NPr Phosphoenolpyruvate-protein phosphotransferase PtsP Guinier plot ln 2.83×100 Rg: 2.3 nm 0 (2.3 nm)-2 s2
(sRg)2I(s)/I(0)
Phosphocarrier protein NPr Phosphoenolpyruvate-protein phosphotransferase PtsP Kratky plot 1.104 0 √3 sRg
p(r)
Phosphocarrier protein NPr Phosphoenolpyruvate-protein phosphotransferase PtsP pair distance distribution function Rg: 2.4 nm 0 Dmax: 8.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Phosphocarrier protein NPr Phosphoenolpyruvate-protein phosphotransferase PtsP PDB (PROTEIN DATA BANK) model
SAXS data from solutions of Enzyme I-Ntr (residues 170-424) complexed with NPr (residues 1-85) in 10 mM Tris 150 mM NaCl 0.5 mM EDTA, pH 7.5 were collected using an in house X-ray source at the NIDDK, National Institutes of Health, Bethesda, MD using a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4π sin θ/λ, and 2θ is the scattering angle). One solute concentration of 3.76 mg/ml was measured. Eight successive 1800 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged and the scattering of the solvent-blank was subtracted. The low angle data collected were obtained from a single concentration scattering curve.

Sample detector distance = UNKNOWN

Phosphocarrier protein NPr (NPr)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   9.3 kDa
 
UniProt   P0A9N0 (1-85)
Sequence   FASTA
 
Phosphoenolpyruvate-protein phosphotransferase PtsP (Enzyme I-Ntr)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   28.4 kDa
 
UniProt   P37177 (170-424)
Sequence   FASTA
 
PDB ID   5T1O