Anomeric Selectivity of Trehalose Transferase with Rare L-Sugars

Mestrom L, Marsden S, van der Eijk H, Laustsen J, Jeffries C, Svergun D, Hagedoorn P, Bento I, Hanefeld U, ACS Catalysis (2020) DOI

SASDHK9 – Trehalose transferase (TreT) from Thermoproteus uzoniensis - with magnesium and UDP-glucose

Trehalose transferase (Trehalose phosphorylase/synthase)
MWexperimental 47 kDa
MWexpected 47 kDa
VPorod 73 nm3
log I(s) 1.09×104 1.09×103 1.09×102 1.09×101
Trehalose transferase (Trehalose phosphorylase/synthase) small angle scattering data  s, nm-1
ln I(s)
Trehalose transferase (Trehalose phosphorylase/synthase) Guinier plot ln 1.09×104 Rg: 2.4 nm 0 (2.4 nm)-2 s2
(sRg)2I(s)/I(0)
Trehalose transferase (Trehalose phosphorylase/synthase) Kratky plot 1.104 0 3 sRg
p(r)
Trehalose transferase (Trehalose phosphorylase/synthase) pair distance distribution function Rg: 2.4 nm 0 Dmax: 8.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Trehalose transferase (Trehalose phosphorylase/synthase) DAMMIN model
Trehalose transferase (Trehalose phosphorylase/synthase) DAMFILT model

log I(s)
 s, nm-1
Trehalose transferase (Trehalose phosphorylase/synthase) GASBOR model

log I(s)
 s, nm-1
Trehalose transferase (Trehalose phosphorylase/synthase) SREFLEX model

Synchrotron SAXS data from solutions of TreT in 50 mM HEPES, 100 NaCl, 4 mM DTT, 20 mM MgCl2, 1 mM UDP-glucose, pH 7 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 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 40 μl sample at 6 mg/ml was injected at a 0.30 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 29 successive 0.500 second frames were collected through the sample elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Experimental molecular weight (MW) assessed using concentration-independent Bayesian inference (Hajizadeh et al., Sci. Reports (2018) 8:7204). MW credibility interval 44-48 kDa. DAMMIN, GASBOR and SREFLEX results are made available in the full entry zip archive.

Trehalose transferase (Trehalose phosphorylase/synthase) (TreT)
Mol. type   Protein
Organism   Thermoproteus uzoniensis
Olig. state   Monomer
Mon. MW   46.7 kDa
 
UniProt   F2L613 (1-401)
Sequence   FASTA