Structural and functional insights into peptidoglycan access for the lytic amidase LytA of Streptococcus pneumoniae.

Mellroth P, Sandalova T, Kikhney A, Vilaplana F, Hesek D, Lee M, Mobashery S, Normark S, Svergun D, Henriques-Normark B, Achour A, MBio 5(1):e01120-13 (2014) PubMed

SASDBG4 – Prophage LytA with choline

Prophage Lytic Amidase with choline
MWI(0) 66 kDa
MWexpected 73 kDa
VPorod 130 nm3
log I(s) 9.84×101 9.84×100 9.84×10-1 9.84×10-2
Prophage Lytic Amidase with choline small angle scattering data  s, nm-1
ln I(s)
Prophage Lytic Amidase with choline Guinier plot ln 9.84×101 Rg: 5.9 nm 0 (5.9 nm)-2 s2
(sRg)2I(s)/I(0)
Prophage Lytic Amidase with choline Kratky plot 1.104 0 3 sRg
p(r)
Prophage Lytic Amidase with choline pair distance distribution function Rg: 5.4 nm 0 Dmax: 17 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Prophage Lytic Amidase with choline SASREF model

Synchrotron SAXS data from solutions of Prophage LytA with choline in 20 mM Tris 150 mM NaCl 5 mM choline chloride 1 ┬ÁM ZnCl2, pH 7.5 were collected on the EMBL X33 camera at the DORIS III storage ring (Hamburg, Germany) using a Pilatus 1M-W detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.15 nm , the range of momentum transfer 0.070 < s < 6.000 nm-1 was covered (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 2 and 10 mg/ml were measured at 10°C. Eight successive 15 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. 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.

SAXS data demonstrate that the wild type (SASDBF4) and bacteriophage LytA are similar in shape and that both form dimers in solution in the presence of choline.

Prophage Lytic Amidase with choline (p-LytA)
Mol. type   Protein
Organism   Streptococcus pneumoniae
Olig. state   Dimer
Mon. MW   36.4 kDa