Structural Dynamics Control Allosteric Activation of Cytohesin Family Arf GTPase Exchange Factors.

Malaby AW, Das S, Chakravarthy S, Irving TC, Bilsel O, Lambright DG, Structure 26(1):106-117.e6 (2018) Europe PMC

SASDCQ7 – Truncated monomeric Cytohesin-3 (Grp1; amino acids 63-399) E161A Arf6 Q67L fusion protein

Grp1 63-399 E161A Arf6 Q67L fusion protein
MWexperimental 58 kDa
MWexpected 60 kDa
VPorod 86 nm3
log I(s) 1.01×100 1.01×10-1 1.01×10-2 1.01×10-3
Grp1 63-399 E161A Arf6 Q67L fusion protein small angle scattering data  s, nm-1
ln I(s)
Grp1 63-399 E161A Arf6 Q67L fusion protein Guinier plot ln 1.01×100 Rg: 3.1 nm 0 (3.1 nm)-2 s2
(sRg)2I(s)/I(0)
Grp1 63-399 E161A Arf6 Q67L fusion protein Kratky plot 1.104 0 √3 sRg
p(r)
Grp1 63-399 E161A Arf6 Q67L fusion protein pair distance distribution function Rg: 3.1 nm 0 Dmax: 11.6 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Truncated monomeric Cytohesin-3 (Grp1; amino acids 63-399) E161A Arf6 Q67L fusion protein Rg histogram Rg, nm
Grp1 63-399 E161A Arf6 Q67L fusion protein EOM/RANCH model
Grp1 63-399 E161A Arf6 Q67L fusion protein EOM/RANCH model
log I(s)
 s, nm-1
Grp1 63-399 E161A Arf6 Q67L fusion protein CORAL model
log I(s)
 s, nm-1
Grp1 63-399 E161A Arf6 Q67L fusion protein OTHER model
Grp1 63-399 E161A Arf6 Q67L fusion protein OTHER model
This experimental profile was derived from SEC-SAXS data collected at the BioCAT Sector 18-ID beamline at the Argonne National Laboratory Advanced Photon Source using a MAR 165 CCD detector. The protein was equilibrated with a 1.2 molar excess of inositol 1,3,4,5-tetrakis phosphate (IP4), concentrated to 10 mg/ml, and injected in 0.1 ml onto a 3 ml Superdex-200 Increase column equilibrated with 20 mM Tris, pH 8.0, 150 mM NaCl, 2 mM MgCl2, 0.1% 2-mercaptoethanol, 5% glycerol, and 0.001 ml IP4. SAXS data sets were acquired with 1 s exposures at 5 s intervals during elution at a flow rate of 0.25 ml/min. Raw SAXS images were radially averaged on a log scale over the q range 0.00621-0.333 Å-1 and normalized by the incident beam intensity. The protein scattering profile was reconstructed by singular value decomposition and linear combination (SVD-LC) as described in Malaby et al. (2015) Methods for analysis of size-exclusion chromatography-small angle X-ray scattering and reconstruction of protein scattering. J Appl Crystallogr 48: 1102-1113.

Storage temperature = UNKNOWN. Concentration min = UNKNOWN

Grp1 63-399 E161A Arf6 Q67L fusion protein
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   60.1 kDa
Sequence   FASTA
 
PDB ID   2R09
 
PDB ID   2R09
 
PDB ID   2R09
 
PDB ID   2R09
 
PDB ID   2R09