Tandem engagement of phosphotyrosines by the dual SH2 domains of p120RasGAP.

Stiegler AL, Vish KJ, Boggon TJ, Structure (2022) Europe PMC

SASDPM4 – Apo SH2-SH3-SH2 domains from p120RasGAP

Ras GTPase-activating protein 1
MWexperimental 34 kDa
MWexpected 101 kDa
VPorod 41 nm3
log I(s) 1.35×10-2 1.35×10-3 1.35×10-4 1.35×10-5
Ras GTPase-activating protein 1 small angle scattering data  s, nm-1
ln I(s)
Ras GTPase-activating protein 1 Guinier plot ln 1.36×10-2 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Ras GTPase-activating protein 1 Kratky plot 1.104 0 √3 sRg
p(r)
Ras GTPase-activating protein 1 pair distance distribution function Rg: 2.7 nm 0 Dmax: 10.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Ras GTPase-activating protein 1 DAMMIF model
log I(s)
 s, nm-1
On the day of data collection the samples were thawed and spun down. 1 mM DTT was added to the samples and buffer. The samples were injected onto a GE Superdex 200 Increase column at 22˚C in buffer containing 20 mM Tris pH 8, 350 mM NaCl, 1 mM DTT at a flow rate of 0.5 mL/min. Data were collected at APS’s BioCAT Beamline18ID using synchrotron radiation from Undulator A. As the sample came off of the column it was first detected by UV, followed by Wyatt DAWN HELEOS II MALS+DLS and Wyatt Optilab T-rEX dRI detectors for measuring molecular weight through MALS-DLS-RI analysis. Following MALS-DLS, the sample flowed to the SAXS sample chamber where data were collected by a Pilatus3 X 1M detector with a wavelength of 1.033 Å, camera length of 3.628 m, an exposure time of 0.5 s, and a q-measurement range of 0.0045 to 0.35 Å-1. Data were normalized using an active beamstop containing a silicon PIN diode.

Ras GTPase-activating protein 1 (p120RasGAP)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   100.8 kDa
 
UniProt   P20936 (174-1047)
Sequence   FASTA