Identification and characterization of MtbNei2-SSB forms novel pre-replicative BER complex in Mycobacterium tuberculosis.

Jyoti Vishwakarma.

SASDPE3 – GST-Endonuclease 8 2/Single-stranded DNA-binding protein (Nei2-SSB) complex

Endonuclease 8 2
Single-stranded DNA-binding protein
MWexperimental 290 kDa
MWexpected 247 kDa
VPorod 734 nm3
log I(s) 3.14×105 3.14×104 3.14×103 3.14×102
Endonuclease 8 2 Single-stranded DNA-binding protein small angle scattering data  s, nm-1
ln I(s)
Endonuclease 8 2 Single-stranded DNA-binding protein Guinier plot ln 3.14×105 Rg: 5.5 nm 0 (5.5 nm)-2 s2
(sRg)2I(s)/I(0)
Endonuclease 8 2 Single-stranded DNA-binding protein Kratky plot 1.104 0 √3 sRg
p(r)
Endonuclease 8 2 Single-stranded DNA-binding protein pair distance distribution function Rg: 6.2 nm 0 Dmax: 21 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Endonuclease 8 2 Single-stranded DNA-binding protein DAMMIF model
SAXS data from solutions of of the Nei2-SSB complex in 20 mM Tris, 100 mM NaCl, pH 8 were collected using an Anton Paar SAXSpace instrument at the CSIR-Central Drug Research Institute (Lucknow, India) equipped with a Mythen2 R 1K detector at a sample-detector distance of 0.317 m and at a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.00 mg/ml was measured at 10°C. Two successive 1800 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.

Endonuclease 8 2 (NEI2-GST fusion)
Mol. type   Protein
Organism   Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Olig. state   Dimer
Mon. MW   54.0 kDa
 
UniProt   P9WNC1 (1-255)
Sequence   FASTA
 
Single-stranded DNA-binding protein (SSB)
Mol. type   Protein
Organism   Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Olig. state   Octamer
Mon. MW   17.4 kDa
 
UniProt   P9WGD5 (1-164)
Sequence   FASTA