Structural basis for SHOC2 modulation of RAS signalling.

Liau NPD, Johnson MC, Izadi S, Gerosa L, Hammel M Bruning JM, Wendorff TJ, Phung W, Hymowitz SG, Sudhamsu J, Nature (2022) Europe PMC

SASDME5 – Leucine-rich repeat protein SHOC-2 bound to serine/threonine-protein phosphatase PP1-gamma catalytic subunit (PP1C) and Ras-related protein M-Ras

Leucine-rich repeat protein SHOC-2
Serine/threonine-protein phosphatase PP1-gamma catalytic subunit
Ras-related protein M-Ras
MWexperimental 125 kDa
MWexpected 126 kDa
VPorod 215 nm3
log I(s) 5.21×101 5.21×100 5.21×10-1 5.21×10-2
Leucine-rich repeat protein SHOC-2 Serine/threonine-protein phosphatase PP1-gamma catalytic subunit Ras-related protein M-Ras small angle scattering data  s, nm-1
ln I(s)
Leucine-rich repeat protein SHOC-2 Serine/threonine-protein phosphatase PP1-gamma catalytic subunit Ras-related protein M-Ras Guinier plot ln 5.22×101 Rg: 3.5 nm 0 (3.5 nm)-2 s2
(sRg)2I(s)/I(0)
Leucine-rich repeat protein SHOC-2 Serine/threonine-protein phosphatase PP1-gamma catalytic subunit Ras-related protein M-Ras Kratky plot 1.104 0 3 sRg
p(r)
Leucine-rich repeat protein SHOC-2 Serine/threonine-protein phosphatase PP1-gamma catalytic subunit Ras-related protein M-Ras pair distance distribution function Rg: 3.6 nm 0 Dmax: 12.4 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Leucine-rich repeat protein SHOC-2 Serine/threonine-protein phosphatase PP1-gamma catalytic subunit Ras-related protein M-Ras BILBOMD model

Synchrotron SAXS data from solutions of SHOC-2-PP1C-M-Ras in 25 mM Tris pH 7.5, 100 mM NaCl, 1 mM TCEP, 1 mM MnCl2, pH 7.5 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS; Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 2.1 m and at a wavelength of λ = 0.1127 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 5.00 μl sample at 60 mg/ml was injected at a 0.50 ml/min flow rate onto a Shodex KW-800 series column at 20°C. 600 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.

Leucine-rich repeat protein SHOC-2
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   65.1 kDa
 
UniProt   Q9UQ13 (2-582)
Sequence   FASTA
 
Serine/threonine-protein phosphatase PP1-gamma catalytic subunit (PP1C)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   37.1 kDa
 
UniProt   P36873 (1-323)
Sequence   FASTA
 
Ras-related protein M-Ras (MRAS)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   24.0 kDa
 
UniProt   O14807 (1-208)
Sequence   FASTA