The Ruminococcus bromii amylosome protein Sas6 binds single and double helical α-glucan structures in starch.

Photenhauer AL, Villafuerte-Vega RC, Cerqueira FM, Armbruster KM, Mareček F, Chen T, Wawrzak Z, Hopkins JB, Vander Kooi CW, Janeček Š, Ruotolo BT, Koropatkin NM, Nat Struct Mol Biol (2024) Europe PMC

SASDPE2 – Starch adherence system protein 6 (Sas6)

Dockerin type I repeat
MWexperimental 63 kDa
MWexpected 69 kDa
VPorod 97 nm3
log I(s) 3.59×10-2 3.59×10-3 3.59×10-4 3.59×10-5
Dockerin type I repeat small angle scattering data  s, nm-1
ln I(s)
Dockerin type I repeat Guinier plot ln 3.59×10-2 Rg: 3.0 nm 0 (3.0 nm)-2 s2
(sRg)2I(s)/I(0)
Dockerin type I repeat Kratky plot 1.104 0 √3 sRg
p(r)
Dockerin type I repeat pair distance distribution function Rg: 2.9 nm 0 Dmax: 9 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Dockerin type I repeat MULTIFOXS model
Synchrotron SAXS data from solutions of starch adherence system protein 6 (Sas6) in phosphate buffered saline, 1 mM TCEP, pH 7 were collected on the BioCAT 18ID beam line at the Advanced Photon Source (APS), Argonne National Laboratory (Lemont, IL, USA) using a Pilatus3 X 1M detector at a sample-detector distance of 3.6 m and at a wavelength of λ = 0.1033 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 150.00 μl sample at 3.5 mg/ml was injected at a 0.60 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 2000 successive 0.500 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Dockerin type I repeat (Sas6)
Mol. type   Protein
Organism   Ruminococcus bromii
Olig. state   Monomer
Mon. MW   68.9 kDa
 
UniProt   A0A2N0UYM2 (31-665)
Sequence   FASTA