Crystallization and preliminary X-ray crystallographic analysis of native and selenomethionyl recombinant tabtoxin-resistance protein complexed with acetyl-coenzyme A.
Tabtoxin-resistance protein (TTR), an acetyltransferase from Pseudomonas syringae pv. tabaci, was overexpressed in Eschericha coli M15 and the TTR fusion protein complexed with acetyl-coenzyme A (AcCoA) was purified and crystallized. Diffraction data were collected to 3.0 A resolution in-house and the crystal was found to belong to space group P2(1), with unit-cell parameters a = 47.6, b = 66.6, c = 53.5 A, beta = 104.3 degrees. Furthermore, a selenomethionine (SeMet) TTR fusion protein derivative was overexpressed in the same expression system and its complex with AcCoA was purified in a reductive environment. The SeMet TTR derivative crystallized in two forms: the first was identical to that observed for native crystals and the second belonged to space group C2, with unit-cell parameters a = 101.7, b = 45.6, c = 84.2 A, beta = 105.8 degrees. Data from the P2(1) crystal form were collected in-house to 2.3 A resolution. Subsequently, three different wavelength data sets of the C2 crystal form to 1.55 A resolution were collected at the Advanced Photon Source at Argonne National Laboratory.
He
H
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Laboratory of Structural Biology, School of Life Science and Engineering, Tsinghua University, Beijing 100084, People's Republic of China.
Ding
Y
Y
Cao
Z
Z
Shao
Y
Y
Bartlam
M
M
Tang
H
H
Jiang
F
F
Liu
Y
Y
Liu
J
J
Zhao
N
N
Rao
Z
Z
D000105:Acetyl Coenzyme A / Q000737:chemistry*; D000123:Acetyltransferases / Q000737:chemistry*; D001426:Bacterial Proteins; D003460:Crystallization; D018360:Crystallography, X-Ray; D011487:Protein Conformation; D011549:Pseudomonas / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry; D012645:Selenomethionine / Q000737:chemistry
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000105:Acetyl Coenzyme A; D012645:Selenomethionine; D000123:Acetyltransferases; C441636:tabtoxin-resistance protein, Pseudomonas syringae