Conformational characterization of full-length X-chromosome-linked inhibitor of apoptosis protein (XIAP) through an integrated approach

Polykretis P, Luchinat E, Bonucci A, Giachetti A, Graewert M, Svergun D, Banci L, IUCrJ 6(5) (2019) DOI

SASDF24 – Full-length X-chromosome linked inhibitor of apoptosis protein (XIAP)

E3 ubiquitin-protein ligase XIAP
MWexperimental 115 kDa
MWexpected 113 kDa
VPorod 207 nm3
log I(s) 1.64×104 1.64×103 1.64×102 1.64×101
E3 ubiquitin-protein ligase XIAP small angle scattering data  s, nm-1
ln I(s)
E3 ubiquitin-protein ligase XIAP Guinier plot ln 1.64×104 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
E3 ubiquitin-protein ligase XIAP Kratky plot 1.104 0 3 sRg
p(r)
E3 ubiquitin-protein ligase XIAP pair distance distribution function Rg: 4.0 nm 0 Dmax: 12.8 nm

Data validation


Fits and models


log I(s)
 s, nm-1
E3 ubiquitin-protein ligase XIAP HADDOCK model

Synchrotron SAXS data from solutions of X-chromosome linked inhibitor of apoptosis protein (XIAP) in Xiap buffer, pH 7.4 were collected using size-exclusion chromatography SAXS (SEC-SAXS) on the EMBL P12 beam line at PETRA III (Hamburg, Germany) using a Pilatus 2M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). SEC-SAXS was performed at 20 °C using the following parameters: Column: Superdex 200 10/300 GL (GE healthcare Life Sciences); Flow rate: 0.5 mL/min; Sample injection concentration: 7.5 mg/mL; Injection volume: 100 μL. The data were collected through the SEC peak of the protein as a series of 15 x 1 second exposures. Each unsubtracted data frame was normalised to the intensity of the transmitted beam and radially averaged and the scattering of an appropriate solvent-blank was subtracted. The resulting subtracted frames were scaled and averaged to generate the final SAXS profile displayed in this entry.

E3 ubiquitin-protein ligase XIAP
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   56.7 kDa
 
UniProt   P98170 (1-497)
Sequence   FASTA