Structure and mechanism of a phage-encoded SAM lyase revises catalytic function of enzyme family.

Guo X, Söderholm A, Kanchugal P S, Isaksen GV, Warsi O, Eckhard U, Trigüis S, Gogoll A, Jerlström-Hultqvist J, Åqvist J, Andersson DI, Selmer M, Elife 10 (2021) Europe PMC

SASDJ65 – S-adenosylmethionine (SAM) lyase Svi3-3 in presence of SAM

Phage-encoded SAM lyase Svi3-3 (including N-terminal His6-tag and Tev cleavage site)
MWexperimental 46 kDa
MWexpected 56 kDa
VPorod 89 nm3
log I(s) 1.18×10-1 1.18×10-2 1.18×10-3 1.18×10-4
Phage-encoded SAM lyase Svi3-3 (including N-terminal His6-tag and Tev cleavage site) small angle scattering data  s, nm-1
ln I(s)
Phage-encoded SAM lyase Svi3-3 (including N-terminal His6-tag and Tev cleavage site) Guinier plot ln 1.19×10-1 Rg: 2.5 nm 0 (2.5 nm)-2 s2
(sRg)2I(s)/I(0)
Phage-encoded SAM lyase Svi3-3 (including N-terminal His6-tag and Tev cleavage site) Kratky plot 1.104 0 √3 sRg
p(r)
Phage-encoded SAM lyase Svi3-3 (including N-terminal His6-tag and Tev cleavage site) pair distance distribution function Rg: 2.5 nm 0 Dmax: 9.1 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of SAM lyase Svi3-3 in presence of SAM in 25 mM Tris-HCl, 150 mM NaCl, 5 mM SAM, pH 8 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Pilatus 2M detector at a sample-detector distance of 4.0 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 10 mg/ml was injected at a 0.16 ml/min flow rate onto a Shodex KW403 column at 25°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted from the SEC elution frames.

Exposure time = UNKNOWN

Phage-encoded SAM lyase Svi3-3 (including N-terminal His6-tag and Tev cleavage site) (Svi3-3)
Mol. type   Protein
Organism   Unknown environmental phage
Olig. state   Trimer
Mon. MW   18.6 kDa
Sequence   FASTA