Structural Characterization of Full-Length Human Dehydrodolichyl Diphosphate Synthase Using an Integrative Computational and Experimental Approach.

Lisnyansky Bar-El M, Lee SY, Ki AY, Kapelushnik N, Loewenstein A, Chung KY, Schneidman-Duhovny D, Giladi M, Newman H, Haitin Y, Biomolecules 9(11) (2019) Europe PMC

SASDHX2 – Dehydrodolichyl diphosphate synthase complex subunit DHDDS

Dehydrodolichyl diphosphate synthase complex subunit DHDDS
MWexperimental 78 kDa
MWexpected 78 kDa
VPorod 122 nm3
log I(s) 3.09×100 3.09×10-1 3.09×10-2 3.09×10-3
Dehydrodolichyl diphosphate synthase complex subunit DHDDS small angle scattering data  s, nm-1
ln I(s)
Dehydrodolichyl diphosphate synthase complex subunit DHDDS Guinier plot ln 3.09×100 Rg: 3.2 nm 0 (3.2 nm)-2 s2
(sRg)2I(s)/I(0)
Dehydrodolichyl diphosphate synthase complex subunit DHDDS Kratky plot 1.104 0 3 sRg
Dmax: 13 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Dehydrodolichyl diphosphate synthase complex subunit DHDDS MODELLER model

Synchrotron SAXS data from solutions of DHDDS in Tris-HCl, 150 mM NaCl, 20 mM 2-mercaptoethanol and 0.02% Triton-X100, pH 7.5 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 8 mg/ml was injected onto a GE Superdex 200 Increase 10/300 column at 20°C. 2000 successive 1 second frames were collected through the SEC-elution profile. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted from the scattering data measured from the sample elution peak.

Dehydrodolichyl diphosphate synthase complex subunit DHDDS (DHDDS)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   39.1 kDa
 
UniProt   Q86SQ9 (1-333)
Sequence   FASTA