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