Structural characterization of bacterial collagenases

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

SASDCL2 – Tandem CBD from Clostridium histolyticum ColG collagenase at pCa 3

Class1 collagenase
MWexperimental 27 kDa
MWexpected 27 kDa
VPorod 36 nm3
log I(s) 9.71×101 9.71×100 9.71×10-1 9.71×10-2
Class1 collagenase small angle scattering data  s, nm-1
ln I(s)
Class1 collagenase Guinier plot ln 9.71×101 Rg: 2.4 nm 0 (2.4 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: 2.4 nm 0 Dmax: 7.5 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 3 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). Solute concentrations ranging between 2 and 6 mg/ml were measured at 10°C. 15 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 and the different curves were scaled for protein concentration. The low angle data collected at lower concentrations were extrapolated to infinite dilution and merged with the higher concentration data to yield the final composite scattering curve.

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