Neutron scattering maps the higher-order assembly of NADPH-dependent assimilatory sulfite reductase

Daniel Murray.

SASDMM8 – Reconstituted sulfite reductase dodecamer with partially deuterated hemoprotein in 0% D2O

Sulfite reductase [NADPH] hemoprotein beta-component
Sulfite reductase [NADPH] flavoprotein alpha-component (His-tagged)
MWexperimental 839 kDa
MWexpected 821 kDa
log I(s) 2.80×100 2.80×10-1 2.80×10-2 2.80×10-3
Sulfite reductase [NADPH] hemoprotein beta-component Sulfite reductase [NADPH] flavoprotein alpha-component (His-tagged) small angle scattering data  s, nm-1
ln I(s)
Sulfite reductase [NADPH] hemoprotein beta-component Sulfite reductase [NADPH] flavoprotein alpha-component (His-tagged) Guinier plot ln 2.81×100 Rg: 11.2 nm 0 (11.2 nm)-2 s2
(sRg)2I(s)/I(0)
Sulfite reductase [NADPH] hemoprotein beta-component Sulfite reductase [NADPH] flavoprotein alpha-component (His-tagged) Kratky plot 1.104 0 3 sRg
p(r)
Sulfite reductase [NADPH] hemoprotein beta-component Sulfite reductase [NADPH] flavoprotein alpha-component (His-tagged) pair distance distribution function Rg: 10.4 nm 0 Dmax: 30.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1

SANS data from solutions of reconstituted sulfite reductase dodecamer with partially deuterated hemoprotein in 0% D2O, 50 mM KPi, 100 mM NaCl, 1 mM EDTA, pH 7.8 were collected using the EQ-SANS (BL-6) spallation neutron source (Oak Ridge, Tennessee, USA). Three instrument configurations were used: 9 m sample-to-detector distance with 1.5 nm wavelength band, 4 m sample-to-detector distance with 0.6 nm wavelength band, and 1.3 m sample-to-detector distance with 0.4 nm wavelength band. Sample concentration: 4.5 mg/mL. Neutron exposure time: 3 hours / instrument configuration. Sample temperature: 8°C.

Sulfite reductase [NADPH] hemoprotein beta-component (SiRHP)
Mol. type   Protein
Organism   Escherichia coli (strain K12)
Olig. state   Tetramer
Mon. MW   64.0 kDa
 
UniProt   P17846 (1-570)
Sequence   FASTA
 
Sulfite reductase [NADPH] flavoprotein alpha-component (His-tagged) (His-tagged SiRFP)
Mol. type   Protein
Organism   Escherichia coli (strain K12)
Olig. state   Octamer
Mon. MW   70.7 kDa
 
UniProt   P38038 (1-590)
Sequence   FASTA