Proteins involved in sleep homeostasis: Biophysical characterization of INC and its partners

Pirone L, Smaldone G, Esposito C, Balasco N, Petoukhov M, Spilotros A, Svergun D, Di Gaetano S, Vitagliano L, Pedone E, Biochimie 131:106-114 (2016) DOI

SASDLN4 – Insomniac protein (INC) – cullin 3 complex

Cullin 3
Insomniac protein
MWexperimental 270 kDa
MWexpected 270 kDa
log I(s) 3.58×10-2 3.58×10-3 3.58×10-4 3.58×10-5
Cullin 3 Insomniac protein small angle scattering data  s, nm-1
ln I(s)
Cullin 3 Insomniac protein Guinier plot ln 3.58×10-2 Rg: 6.7 nm 0 (6.7 nm)-2 s2
(sRg)2I(s)/I(0)
Cullin 3 Insomniac protein Kratky plot 1.104 0 √3 sRg
p(r)
Cullin 3 Insomniac protein pair distance distribution function Rg: 6.7 nm 0 Dmax: 25.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Cullin 3 Insomniac protein SASREF model
Synchrotron SAXS data from solutions of Insomniac protein (INC) – cullin 3 complex in 50 mM TrisHCl, 300 mM NaCl, 2 mM DTT, pH 8 were collected on the B21 beam line at the Diamond Light Source storage ring (Didcot, UK) using a Pilatus 2M detector 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 6.00 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.

Storage temperature = UNKNOWN. Sample detector distance = UNKNOWN

Cullin 3 (Cul3)
Mol. type   Protein
Organism   Drosophila melanogaster
Olig. state   Pentamer
Mon. MW   12.0 kDa
Sequence   FASTA
 
Insomniac protein (INC)
Mol. type   Protein
Organism   Drosophila melanogaster
Olig. state   Pentamer
Mon. MW   41.9 kDa
Sequence   FASTA