skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Thermostable multicopper oxidase from Thermusthermophilus HB27: crystallization and preliminaryX-ray diffraction analysis of apo and holo forms

Abstract

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 {angstrom} resolution obtained using synchrotron radiation at 100 K are reported. The crystals belonged to space group C222{sub 1}, with unit-cell parameters a = 93.6, b = 110.3, c = 96.3 {angstrom}. A monomer in the asymmetric unit yielded a Matthews coefficient (V{sub M}) of 2.60 {angstrom}{sup 3} Da{sup -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 {angstrom} 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 {angstrom}.

Authors:
; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
USDOE SC OFFICE OF SCIENCE (SC)
OSTI Identifier:
1041613
Report Number(s):
BNL-96849-2012-JA
Journal ID: ISSN 1744-3091; R&D Project: LS001; TRN: US201212%%31
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
Additional Journal Information:
Journal Volume: 67; Journal Issue: 12; Journal ID: ISSN 1744-3091
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; CHLORIDES; CRYSTALLIZATION; DIFFRACTION; ENZYMES; MONOMERS; OXIDASES; RESOLUTION; SOLVENTS; SPACE GROUPS; SYNCHROTRON RADIATION; X-RAY DIFFRACTION; national synchrotron light source

Citation Formats

Serrano-Posada H., Stojanoff V., Valderrama B., and Rudino-Pinera E. Thermostable multicopper oxidase from Thermusthermophilus HB27: crystallization and preliminaryX-ray diffraction analysis of apo and holo forms. United States: N. p., 2011. Web. doi:10.1107/S174430911103805X.
Serrano-Posada H., Stojanoff V., Valderrama B., & Rudino-Pinera E. Thermostable multicopper oxidase from Thermusthermophilus HB27: crystallization and preliminaryX-ray diffraction analysis of apo and holo forms. United States. doi:10.1107/S174430911103805X.
Serrano-Posada H., Stojanoff V., Valderrama B., and Rudino-Pinera E. Sat . "Thermostable multicopper oxidase from Thermusthermophilus HB27: crystallization and preliminaryX-ray diffraction analysis of apo and holo forms". United States. doi:10.1107/S174430911103805X.
@article{osti_1041613,
title = {Thermostable multicopper oxidase from Thermusthermophilus HB27: crystallization and preliminaryX-ray diffraction analysis of apo and holo forms},
author = {Serrano-Posada H. and Stojanoff V. and Valderrama B. and Rudino-Pinera E.},
abstractNote = {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 {angstrom} resolution obtained using synchrotron radiation at 100 K are reported. The crystals belonged to space group C222{sub 1}, with unit-cell parameters a = 93.6, b = 110.3, c = 96.3 {angstrom}. A monomer in the asymmetric unit yielded a Matthews coefficient (V{sub M}) of 2.60 {angstrom}{sup 3} Da{sup -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 {angstrom} 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 {angstrom}.},
doi = {10.1107/S174430911103805X},
journal = {Acta Crystallographica Section F: Structural Biology and Crystallization Communications},
issn = {1744-3091},
number = 12,
volume = 67,
place = {United States},
year = {2011},
month = {9}
}