TY - JOUR
T1 - Purification, crystallization, and preliminary X-ray diffraction analysis of the Tricorn protease hexamer from Thermoplasma acidophilum
AU - Bosch, Jürgen
AU - Tamura, Tomohiro
AU - Bourenkov, Gleb
AU - Baumeister, Wolfgang
AU - Essen, Lars Oliver
PY - 2001
Y1 - 2001
N2 - Tricorn protease from Thermoplasma acidophilum is a hexameric enzyme; in vivo the hexamers assemble further to form large icosahedral capsids of 14.6 MDa. Recombinant Tricorn protease was purified as an enzymatically active hexamer of 0.72 MDa that formed crystals of octahedral morphology under low-ionic-strength conditions. These crystals belong to space group C2 with unit cell dimensions a = 307.5 Å, b = 163.2 Å, c = 220.9 Å, β = 105.5° and diffract to 2.2-Å resolution using high-brilliance synchrotron radiation. Based on analysis of the self-rotation function and the presence of a pseudo-origin peak in the native Patterson map, a packing model was derived for the complex, comprising 1.5 hexamers per asymmetric unit with a solvent content of 43%. Due to the ninefold noncrystallographic symmetry the Tricorn crystals represent an interesting case for phasing X-ray crystallographic data by electron microscopic phase information.
AB - Tricorn protease from Thermoplasma acidophilum is a hexameric enzyme; in vivo the hexamers assemble further to form large icosahedral capsids of 14.6 MDa. Recombinant Tricorn protease was purified as an enzymatically active hexamer of 0.72 MDa that formed crystals of octahedral morphology under low-ionic-strength conditions. These crystals belong to space group C2 with unit cell dimensions a = 307.5 Å, b = 163.2 Å, c = 220.9 Å, β = 105.5° and diffract to 2.2-Å resolution using high-brilliance synchrotron radiation. Based on analysis of the self-rotation function and the presence of a pseudo-origin peak in the native Patterson map, a packing model was derived for the complex, comprising 1.5 hexamers per asymmetric unit with a solvent content of 43%. Due to the ninefold noncrystallographic symmetry the Tricorn crystals represent an interesting case for phasing X-ray crystallographic data by electron microscopic phase information.
KW - Electron microscopy
KW - Self-compartmentalizing protease
KW - Thermophile
KW - Tricorn
KW - X-ray crystallography
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U2 - 10.1006/jsbi.2001.4360
DO - 10.1006/jsbi.2001.4360
M3 - Article
C2 - 11469880
AN - SCOPUS:18744376560
SN - 1047-8477
VL - 134
SP - 83
EP - 87
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 1
ER -