Structural characterization of bacterial collagenases

Bauer R, University of Arkansas PhD thesis 2431 (2017) URL

SASDCP2 – Tandem CBD from Clostridium histolyticum ColG collagenase at pCa 6

Class1 collagenase
MWexperimental 27 kDa
MWexpected 27 kDa
VPorod 33 nm3
log I(s) 2.61×102 2.61×101 2.61×100 2.61×10-1
Class1 collagenase small angle scattering data  s, nm-1
ln I(s)
Class1 collagenase Guinier plot ln 2.62×102 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Class1 collagenase Kratky plot 1.104 0 √3 sRg
p(r)
Class1 collagenase pair distance distribution function Rg: 2.7 nm 0 Dmax: 12.8 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Class1 collagenase DAMMIF model
Synchrotron SAXS data from solutions of the tandem CBD from ColG collagenase at pCa 6 in 10 mM HEPES 100 mM NaCL, 2% glycerol, pH 7.5 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS, Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 1.5 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 at 10°C. The data were normalized to the intensity of the transmitted beam and radially averaged and the scattering of the solvent-blank was subtracted.

X-ray Exposure time = UNKNOWN. Number of frames = UNKNOWN

Class1 collagenase (tandem CBD)
Mol. type   Protein
Organism   Hathewaya histolytica
Olig. state   Monomer
Mon. MW   27.1 kDa
 
UniProt   Q9X721 (883-1118)
Sequence   FASTA