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

SASDCM7 – Truncated monomeric Cytohesin-3 (Grp1; amino acids 63-399) E161A 6GS Arf6 Q67L fusion protein

Grp1 63-399 E161A 6GS Arf6 Q67L fusion protein
MWexperimental 62 kDa
MWexpected 61 kDa
VPorod 93 nm3
log I(s) 1.02×100 1.02×10-1 1.02×10-2 1.02×10-3
Grp1 63-399 E161A 6GS Arf6 Q67L fusion protein small angle scattering data  s, nm-1
ln I(s)
Grp1 63-399 E161A 6GS Arf6 Q67L fusion protein Guinier plot ln 1.02×100 Rg: 3.3 nm 0 (3.3 nm)-2 s2
(sRg)2I(s)/I(0)
Grp1 63-399 E161A 6GS Arf6 Q67L fusion protein Kratky plot 1.104 0 3 sRg
p(r)
Grp1 63-399 E161A 6GS Arf6 Q67L fusion protein pair distance distribution function Rg: 3.3 nm 0 Dmax: 11.9 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Grp1 63-399 E161A 6GS Arf6 Q67L fusion protein DAMMIF model
Grp1 63-399 E161A 6GS Arf6 Q67L fusion protein DAMFILT 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.

Grp1 63-399 E161A 6GS Arf6 Q67L fusion protein
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   61.0 kDa
Sequence   FASTA