DacA, GlmM and the DacA/GlmM complex

Tommaso Tosi.

SASDE98 – Solution structure of Diadenylate cyclase (DacA) from Staphylococcus aureus

Diadenylate cyclase
MWexperimental 40 kDa
MWexpected 39 kDa
VPorod 57 nm3
log I(s) 3.14×10-2 3.14×10-3 3.14×10-4 3.14×10-5
Diadenylate cyclase small angle scattering data  s, nm-1
ln I(s)
Diadenylate cyclase Guinier plot ln 3.14×10-2 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Diadenylate cyclase Kratky plot 1.104 0 3 sRg
p(r)
Diadenylate cyclase pair distance distribution function Rg: 2.6 nm 0 Dmax: 8.6 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Diadenylate cyclase DAMFILT model

Synchrotron SAXS data from solutions of the DacA in 30 mM Tris, 150 mM NaCl, pH 7.5 were collected using size-exclusion chromatography SAXS (SEC-SAXS) on the B21 beam line at the Diamond Light Source (Oxfordshire, UK) using a Pilatus 2M detector at a sample-to-detector distance of 4.014 m and X-ray wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). SEC-SAXS was performed at 20°C using the following parameters: Column: Superdex 200 5/150 (GE Healthcare); Flow rate: 0.075 mL/min; Sample injection concentration: 12.0 mg/mL; Injection volume: 45 μL. The data obtained through the sample elution peak (collected as consecutive 3 s exposures) were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted and the individual subtracted data sets were scaled and averaged to generate the scattering profile displayed in this entry.

Diadenylate cyclase (DacA)
Mol. type   Protein
Organism   Staphylococcus aureus (strain USA300)
Olig. state   Dimer
Mon. MW   19.4 kDa
 
UniProt   A0A0H2XIU4 (100-269)
Sequence   FASTA