TY - JOUR
T1 - Expression, purification, crystallization and preliminary X-ray analysis of phosphotransacetylase from Methanosarcina thermophila
AU - Iyer, Prabha P.
AU - Lawrence, Sarah H.
AU - Yennawar, Hemant P.
AU - Ferry, James G.
PY - 2003/8/1
Y1 - 2003/8/1
N2 - Phosphotransacetylase (Pta) from the anaerobic archaeon Methanosarcina thermophila has been heterologously expressed in a soluble form which facilitated crystallization using the hanging-drop vapor-diffusion method with ammonium sulfate as a precipitant. This is the first report of the crystallization of any Pta. While the M. thermophila Pta has high sequence identity to Ptas from other organisms, it has no homology to any previously crystallized proteins. The protein crystallized in space group I41, with unit-cell parameters a = b = 114.8, c = 127.8 Å, α = β = γ = 90°. The crystals diffracted to 2.5 Å resolution using Cu Kα radiation. The enzyme had previously been reported to exist as a monomer; however, the self-rotation function showed the presence of a non-crystallographic symmetry axis at ψ = 90, φ = 90, κ = 180°, suggesting oligomerization. Dynamic light-scattering analysis supported a dimeric state for Pta in solution.
AB - Phosphotransacetylase (Pta) from the anaerobic archaeon Methanosarcina thermophila has been heterologously expressed in a soluble form which facilitated crystallization using the hanging-drop vapor-diffusion method with ammonium sulfate as a precipitant. This is the first report of the crystallization of any Pta. While the M. thermophila Pta has high sequence identity to Ptas from other organisms, it has no homology to any previously crystallized proteins. The protein crystallized in space group I41, with unit-cell parameters a = b = 114.8, c = 127.8 Å, α = β = γ = 90°. The crystals diffracted to 2.5 Å resolution using Cu Kα radiation. The enzyme had previously been reported to exist as a monomer; however, the self-rotation function showed the presence of a non-crystallographic symmetry axis at ψ = 90, φ = 90, κ = 180°, suggesting oligomerization. Dynamic light-scattering analysis supported a dimeric state for Pta in solution.
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U2 - 10.1107/S0907444903013428
DO - 10.1107/S0907444903013428
M3 - Article
C2 - 12876371
AN - SCOPUS:0043069556
SN - 0907-4449
VL - 59
SP - 1517
EP - 1520
JO - Acta Crystallographica Section D: Biological Crystallography
JF - Acta Crystallographica Section D: Biological Crystallography
IS - 8
ER -