Redox Modulation of Oligomeric State in Proline Utilization A.

Korasick DA, Campbell AC, Christgen SL, Chakravarthy S, White TA, Becker DF, Tanner JJ, Biophys J 114(12):2833-2843 (2018) PubMed

SASDDQ3 – Proline utilization A from Bradyrhizobium diazoefficiens (formerly Bradyrhizobium japonicum) collected by SEC-SAXS

Bifunctional protein PutA
MWexperimental 434 kDa
MWexpected 430 kDa
VPorod 582 nm3
log I(s) 4.61×102 4.61×101 4.61×100 4.61×10-1
Bifunctional protein PutA small angle scattering data  s, nm-1
ln I(s)
Bifunctional protein PutA Guinier plot ln 4.61×102 Rg: 5.2 nm 0 (5.2 nm)-2 s2
(sRg)2I(s)/I(0)
Bifunctional protein PutA Kratky plot 1.104 0 3 sRg
p(r)
Bifunctional protein PutA pair distance distribution function Rg: 5.2 nm 0 Dmax: 14.2 nm

Experimental data validation


Fits and models


log I(s)
 s, nm-1
Bifunctional protein PutA PDB model
Synchrotron SAXS data from solutions of proline utilization A from Bradyrhizobium diazoefficiens (formerly Bradyrhizobium japonicum) were collected using SEC-SAXS in 50 mM Tris, 50 mM NaCl, 0.5 mM TCEP, 5% (v/v) glycerol, pH 7.8 on the BioCAT 18ID beam line at the Advanced Photon Source (APS, Argonne, IL, USA) using a Pilatus 100K detector at a sample-detector distance of 3.5 m and at a wavelength of λ = 0.103 nm (l(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). The SEC-SAXS elution was collected at 22°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Bifunctional protein PutA (BjPutA)
Mol. type   Protein
Organism   Bradyrhizobium diazoefficiens (strain JCM 10833 / IAM 13628 / NBRC 14792 / USDA 110)
Olig. state   Tetramer
Mon. MW   107.6 kDa
 
UniProt   Q89E26
Sequence   FASTA
 
PDB code   3HAZ