Phase separation of a microtubule plus-end tracking protein into a fluid fractal network

Czub M, Uliana F, Grubić T, Padeste C, Rosowski K, Dufresne E, Menzel A, Vakonakis I, Gasser U, Steinmetz M, (2024) DOI

SASDUU6 – Nuclear fusion protein BIK1 coiled-coil in a high salt buffer

Nuclear fusion protein BIK1
MWexperimental 65 kDa
MWexpected 55 kDa
VPorod 282 nm3
log I(s) 4.22×10-2 4.22×10-3 4.22×10-4 4.22×10-5
Nuclear fusion protein BIK1 small angle scattering data  s, nm-1
ln I(s)
Nuclear fusion protein BIK1 Guinier plot ln 4.23×10-2 Rg: 8 nm 0 (8 nm)-2 s2
(sRg)2I(s)/I(0)
Nuclear fusion protein BIK1 Kratky plot 1.104 0 √3 sRg
p(r)
Nuclear fusion protein BIK1 pair distance distribution function Rg: 8.0 nm 0 Dmax: 27 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Nuclear fusion protein BIK1 DAMFILT model
Synchrotron SAXS data from solutions of BIK1 coiled-coil in 20 mM Tris-HCl pH 7.5, 500 mM NaCl, 2% glycerol, 1 mM DTT were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.09464 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 6 mg/ml was injected at a 0.16 ml/min flow rate onto a Shodex KW404-4F column at 15°C. 37 successive 3.200 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.

Note: The ab initio model displayed in this entry represents the bead-occupancy and volume-corrected shape obtained from the spatial alignment of several individual DAM models (DAMFILT) and does not reflect the fit to the displayed data.

Nuclear fusion protein BIK1 (Bik1 CC)
Mol. type   Protein
Organism   Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Olig. state   Dimer
Mon. MW   27.7 kDa
 
UniProt   P11709 (182-389)
Sequence   FASTA