A bacteriophage mimic of the bacterial nucleoid-associated protein Fis.

Chakraborti S, Balakrishnan D, Trotter AJ, Gittens WH, Yang AWH, Jolma A, Paterson JR, Świątek S, Plewka J Curtis FA, Bowers LY, Pålsson LO, Hughes TR, Taube M, Kozak M, Heddle JG, Sharples GJ, Biochem J (2020) Europe PMC

SASDHS4 – Protein ninH from Bacteriophage lambda (N-terminal histidine tagged) T15A mutated

Protein ninH
MWexperimental 20 kDa
MWexpected 20 kDa
VPorod 33 nm3
log I(s) 2.53×100 2.53×10-1 2.53×10-2 2.53×10-3
Protein ninH small angle scattering data  s, nm-1
ln I(s)
Protein ninH Guinier plot ln 2.53×100 Rg: 2.1 nm 0 (2.1 nm)-2 s2
(sRg)2I(s)/I(0)
Protein ninH Kratky plot 1.104 0 √3 sRg
p(r)
Protein ninH pair distance distribution function Rg: 2.2 nm 0 Dmax: 8.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Protein ninH MODELLER model
log I(s)
 s, nm-1
Protein ninH DAMFILT model
log I(s)
 s, nm-1
Protein ninH DAMMIF model
SAXS data from solutions of Protein ninH (T15A) in 150 mM NaCl, 50 mM phosphate buffer, pH 7.4, 1 mM EDTA were collected using Xenocs Xeuss 2.0 instrument (Department of Macromolecular Physics, Adam Mickiewicz University, Poznań, Poland) equipped with a MetalJet D2 microfocus X-ray generator and a Pilatus 3R 1M detector at a wavelength of λ = 0.134 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). A sample was collected from a SEC column immediately prior to batch analysis. One solute concentration of 4.00 mg/ml was measured at 22°C. Three successive 600 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged and each exposure checked for radiation damage; the scattering of the solvent-blank was subtracted.

The resulting SAXS data used to validate the model of NinH derived from MODELLER.

Protein ninH (ninH -T15A)
Mol. type   Protein
Organism   Escherichia phage lambda
Olig. state   Dimer
Mon. MW   10.0 kDa
 
UniProt   P03771 (1-68)
Sequence   FASTA