A molecular mechanism for LINC complex branching by structurally diverse SUN-KASH 6:6 assemblies.

Gurusaran M, Davies OR, Elife 10 (2021) Europe PMC

SASDJE5 – LINC complex between the SUN domain of SUN1 and KASH domain of Nesprin-1 - SUN1-KASH1 6:6 complex

SUN domain-containing protein 1
Nesprin-1
MWexperimental 152 kDa
MWexpected 157 kDa
VPorod 254 nm3
log I(s) 1.31×10-1 1.31×10-2 1.31×10-3 1.31×10-4
SUN domain-containing protein 1 Nesprin-1 small angle scattering data  s, nm-1
ln I(s)
SUN domain-containing protein 1 Nesprin-1 Guinier plot ln 1.32×10-1 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
SUN domain-containing protein 1 Nesprin-1 Kratky plot 1.104 0 3 sRg
p(r)
SUN domain-containing protein 1 Nesprin-1 pair distance distribution function Rg: 3.9 nm 0 Dmax: 13 nm

Data validation


Fits and models


log I(s)
 s, nm-1
SUN domain-containing protein 1 Nesprin-1 CORAL model

log I(s)
 s, nm-1
SUN domain-containing protein 1 Nesprin-1 CORAL model

log I(s)
 s, nm-1
SUN domain-containing protein 1 Nesprin-1 DAMMIN model
SUN domain-containing protein 1 Nesprin-1 DAMMIN model

Synchrotron SAXS data from solutions of the Nesprin-1-KASH/SUN1 6:6 complex in 20 mM Tris pH 8.0, 150 mM KCl, were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Pilatus 2M detector at a wavelength of λ = 0.095 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 100.00 μl sample at 19 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 30 successive 3 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.

SUN domain-containing protein 1 (SUN1)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Hexamer
Mon. MW   22.5 kDa
 
UniProt   O94901 (616-812)
Sequence   FASTA
 
Nesprin-1 (SYNE1)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Hexamer
Mon. MW   3.6 kDa
 
UniProt   Q8NF91 (8769-8797)
Sequence   FASTA
 
PDB ID   6R15