Structural insights of the enzymes from the chitin utilization locus of Flavobacterium johnsoniae.

Mazurkewich S, Helland R, Mackenzie A, Eijsink VGH, Pope PB, Brändén G, Larsbrink J, Sci Rep 10(1):13775 (2020) Europe PMC

SASDHU9 – Solution structure of middle domain of the multi-domain GH18 chitinase ChiA from Flavobacterium johnsoniae

Chitinase ChiA
MWexperimental 71 kDa
MWexpected 70 kDa
VPorod 99 nm3
log I(s) 2.84×102 2.84×101 2.84×100 2.84×10-1
Chitinase ChiA small angle scattering data  s, nm-1
ln I(s)
Chitinase ChiA Guinier plot ln 2.85×102 Rg: 4.7 nm 0 (4.7 nm)-2 s2
(sRg)2I(s)/I(0)
Chitinase ChiA Kratky plot 1.104 0 √3 sRg
p(r)
Chitinase ChiA pair distance distribution function Rg: 4.9 nm 0 Dmax: 16.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Chitinase ChiA DAMMIN model
Synchrotron SAXS data from solutions of GH18 chitinase (ChiA) in 50 mM Tris, 250 mM NaCl, 0.25 mM DTT, pH 8 were collected on the BL4-2 beam line at the Stanford Synchrotron Radiation Lightsource (SSRL; Menlo Park, CA, USA) using a Pilatus3 X 1M detector at a sample-detector distance of 1.8 m and at a wavelength of λ = 0.1127 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 5.00 mg/ml was measured. 10 successive 1 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.

Experimental temperature: UNKNOWN.

Chitinase ChiA (ChiA)
Mol. type   Protein
Organism   Flavobacterium johnsoniae
Olig. state   Monomer
Mon. MW   70.4 kDa
 
UniProt   A5FB63 (464-1137)
Sequence   FASTA