Structural, Thermodynamic and Enzymatic Characterization of N,N-Diacetylchitobiose Deacetylase from Pyrococcus chitonophagus.

Biniek-Antosiak K, Bejger M, Śliwiak J, Baranowski D, Mohammed ASA, Svergun DI, Rypniewski W, Int J Mol Sci 23(24) (2022) Europe PMC

SASDMB9 – Diacetylchitobiose deacetylase (oligomeric mixture of hexamers and dodecamers)

Diacetylchitobiose deacetylase

MW^experimental	185	kDa
MW^expected	185	kDa

log I(s) 4.85×10⁴ 4.85×10³ 4.85×10² 4.85×10¹

Diacetylchitobiose deacetylase small angle scattering data

s, nm^-1

Log plot Log-Log plot

ln I(s)

Diacetylchitobiose deacetylase Guinier plot

ln 4.85×10⁴ R_g: 3.9 nm 0 (3.9 nm)^-2 s²

(sR_g)²I(s)/I(0)

Diacetylchitobiose deacetylase Kratky plot

1.104 0 √3 sR_g

Data validation

Fits and models

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

Diacetylchitobiose deacetylase PYMOL model

Synchrotron SAXS data from solutions of diacetylchitobiose deacetylase in 20 mM TRIS, 200 mM NaCl, pH 7.4 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). One solute concentration of 4.70 mg/ml was measured at 20°C. 40 successive 0.095 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.

Approximate volume fractions: 85-90% hexamer; 10-15% dodecamer. Additional concentration series data spanning 0.25 - 3.5 mg/ml and the corresponding hexamer-dodecamer oligomeric volume fractions are provided in the full entry zip archive.

Diacetylchitobiose deacetylase (DAC-74)
Mol. type		Protein
Organism		Thermococcus chitonophagus
Olig. state		Other
Mon. MW		30.9 kDa

UniProt		A0A160VQZ8 (1-267)
Sequence		FASTA

PDB ID		8BGN