Correlation Map, a goodness-of-fit test for one-dimensional X-ray scattering spectra.

Franke D, Jeffries CM, Svergun DI, Nat Methods 12(5):419-22 (2015) Europe PMC

SASDAA6 – Human serum albumin monomer and mixtures

Human serum albumin monomer
MWexperimental 61 kDa
MWexpected 66 kDa
VPorod 103 nm3
log I(s) 5.21×102 5.21×101 5.21×100 5.21×10-1
Human serum albumin monomer small angle scattering data  s, nm-1
ln I(s)
Human serum albumin monomer Guinier plot ln 5.21×102 Rg: 2.8 nm 0 (2.8 nm)-2 s2
(sRg)2I(s)/I(0)
Human serum albumin monomer Kratky plot 1.104 0 3 sRg
p(r)
Human serum albumin monomer pair distance distribution function Rg: 2.8 nm 0 Dmax: 8.4 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Human serum albumin monomer DAMMIF model

log I(s)
 s, nm-1
Human serum albumin monomer PDB model

log I(s)
 s, nm-1
Human serum albumin monomer PDB model

log I(s)
 s, nm-1

log I(s)
 s, nm-1

log I(s)
 s, nm-1

X-ray synchrotron radiation scattering data from solutions of Human serum albumin in 50 mM HEPES, pH 7.5 were collected on the P12 beam line at Petra-III (Hamburg, Germany) using a Pilatus 2M detector (I(s) vs s, where s = 4π sin θ/λ; 2θ is the scattering angle and λ = 0.12 nm). SAXS data of the HSA monomer were obtained from averaging 38 x 1 second frames spanning a separated HSA monomer elution peak from in-line size exclusion chromatography-SAXS (top four profiles and the respective ab initio and atomic model fits). The bottom three profiles are derived from un-purified stock solutions of HSA, consisting of monomers, dimers, and higher oligomeric species at three concentrations: 20 mg/ml, 10 mg/ml and 5 mg/ml. These data are included to illustrate the effects of repulsive interparticle interactions. The atomic model monomer fits to the HSA mixtures (using PDB 1E78_chain_A) are also shown, with chi2 values of 1.99 (20 mg/ml), 1.27 (10 mg/ml) and 1.43 (5 mg/ml). The p-value determined using Correlation map for the monomer fit to the mixture data is 0 in all instances, indicating statistically significant differences are present between the model and data (cf. a chi2 of 1.01 and p-value of 0.035 for the purified monomer model fit to the SEC-SAXS data).

Tags: benchmark
Human serum albumin monomer (HSA)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   66.5 kDa
 
UniProt   P02768 (25-609)
Sequence   FASTA
 
PDB code   1E78
 
PDB code   1E78