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