Quantitative Conformational Analysis of Functionally Important Electrostatic Interactions in the Intrinsically Disordered Region of Delta Subunit of Bacterial RNA Polymerase.

Kuban V Srb P, Stegnerova H, Padrta P, Zachrdla M, Jasenakova Z, Šanderová H, Vítovská D, Krasny L, Koval T, Dohnalek J, Ziemska-Legi Cka J, Grynberg M, Jarnot P, Gruca A, Jensen MR, Blackledge M, Zidek L, J Am Chem Soc (2019) Europe PMC

SASDFC8 – Delta subunit of RNA polymerase, RNAP (B. subtilis): Lysine to glutamate mutant, 800mM NaCl

DNA-directed RNA polymerase subunit delta - mutant
MWexperimental 33 kDa
MWexpected 20 kDa
VPorod 74 nm3
log I(s) 4.20×103 4.20×102 4.20×101 4.20×100
DNA-directed RNA polymerase subunit delta - mutant small angle scattering data  s, nm-1
ln I(s)
DNA-directed RNA polymerase subunit delta - mutant Guinier plot ln 4.20×103 Rg: 4.5 nm 0 (4.5 nm)-2 s2
(sRg)2I(s)/I(0)
DNA-directed RNA polymerase subunit delta - mutant Kratky plot 1.104 0 √3 sRg
Dmax: 21 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of Lys-Glu mutant RNAP in 20 mM Phosphate buffer, 800 mM NaCl, 0.05% NaN3, pH 6.6 were collected on the EMBL P12 beam line at 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). One solute concentration of 2.60 mg/ml was measured at 20°C. 20 successive 0.050 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

DNA-directed RNA polymerase subunit delta - mutant (rpoE KE mutant)
Mol. type   Protein
Organism   Bacillus subtilis
Olig. state   Monomer
Mon. MW   20.4 kDa
 
UniProt   P12464 (1-173)
Sequence   FASTA