Open AccessThermostable multicopper oxidase from Thermus thermophilus HB27: crystallization and preliminary X‐ray diffraction analysis of apo and holo forms
Author(s) -
SerranoPosada Hugo,
Valderrama Brenda,
Stojanoff Vivian,
RudiñoPiñera Enrique
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430911103805x
Subject(s) - thermus thermophilus , crystallization , crystallography , chemistry , solvent , resolution (logic) , materials science , diffraction , organic chemistry , escherichia coli , optics , physics , biochemistry , gene , artificial intelligence , computer science
A thermostable multicopper oxidase from Thermus thermophilus HB27 ( Tth ‐MCO) was successfully crystallized using the sitting‐drop and hanging‐drop vapour‐diffusion methods. Crystallization conditions and preliminary X‐ray diffraction data to 1.5 Å resolution obtained using synchrotron radiation at 100 K are reported. The crystals belonged to space group C 222 1 , with unit‐cell parameters a = 93.6, b = 110.3, c = 96.3 Å. A monomer in the asymmetric unit yielded a Matthews coefficient ( V M ) of 2.60 Å 3 Da −1 and a solvent content of 53%. An inactive enzyme form, apo‐ Tth ‐MCO, was also crystallized and diffraction data were collected to 1.7 Å resolution. In addition, a second inactive form of the enzyme, Hg‐ Tth ‐MCO, was obtained by soaking apo‐ Tth ‐MCO crystals with mercury(II) chloride and data were collected to a resolution of 1.7 Å.