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