TY - JOUR
T1 - Isolation and purification of Thermus thermophilus HpaB by a crystallization approach
AU - Soulimane, Tewfik
AU - Okane, Sarah R.
AU - Kolaj, Olga
PY - 2010
Y1 - 2010
N2 - The oxygenase HpaB is a component of the 4-hydroxyphenylacetate 3-monooxygenase enzyme that is responsible for the hydroxylation of 4-hydroxy-phenylacetate. It utilizes molecular oxygen and a reduced flavin, which is provided by HpaC, the second component of the enzyme. While isolating integral membrane respiratory complexes from Thermus thermophilus, microcrystals of HpaB were formed. Further purification of the enzyme was achieved by repetitive crystallization. Subsequently, well shaped single crystals of the native enzyme that diffract to 1.82 Å resolution were grown in sitting drops. They belong to the orthorhombic space group I222, with unit-cell parameters a = 91.3, b = 99.8, c = 131.7 Å.
AB - The oxygenase HpaB is a component of the 4-hydroxyphenylacetate 3-monooxygenase enzyme that is responsible for the hydroxylation of 4-hydroxy-phenylacetate. It utilizes molecular oxygen and a reduced flavin, which is provided by HpaC, the second component of the enzyme. While isolating integral membrane respiratory complexes from Thermus thermophilus, microcrystals of HpaB were formed. Further purification of the enzyme was achieved by repetitive crystallization. Subsequently, well shaped single crystals of the native enzyme that diffract to 1.82 Å resolution were grown in sitting drops. They belong to the orthorhombic space group I222, with unit-cell parameters a = 91.3, b = 99.8, c = 131.7 Å.
UR - http://www.scopus.com/inward/record.url?scp=77749242841&partnerID=8YFLogxK
U2 - 10.1107/S1744309110003714
DO - 10.1107/S1744309110003714
M3 - Article
C2 - 20208179
AN - SCOPUS:77749242841
SN - 1744-3091
VL - 66
SP - 352
EP - 356
JO - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
JF - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
IS - 3
ER -