Crystallization and X-ray diffraction studies of a two-domain laccase from Streptomyces griseoflavus.
Laccase (EC 1.10.3.2) is one of the most common copper-containing oxidases; it is found in many organisms and catalyzes the oxidation of primarily phenolic compounds by oxygen. Two-domain laccases have unusual thermostability, resistance to inhibitors and an alkaline optimum of activity. The causes of these properties in two-domain laccases are poorly understood. A recombinant two-domain laccase (SgfSL) was cloned from the genome of Streptomyces griseoflavus Ac-993, expressed in Escherichia coli and purified to homogeneity. The crystals of SgfSL belonged to the monoclinic space group P21, with unit-cell parameters a = 74.64, b = 94.72, c = 117.40 Å, β = 90.672°, and diffraction data were collected to 2.0 Å resolution using a synchrotron-radiation source. Two functional trimers per asymmetric unit correspond to a Matthews coefficient of 1.99 Å(3) Da(-1) according to the monomer molecular weight of 35.6 kDa.
Institute of Protein Research, Federal Agency of Scientific Organization, Institutskaya 4, 142290 Pushchino, Moscow Region, Russian Federation.
Svetlana
S
Tishchenko
Institute of Protein Research, Federal Agency of Scientific Organization, Institutskaya 4, 142290 Pushchino, Moscow Region, Russian Federation.
Azat
A
Gabdulkhakov
G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Federal Agency of Scientific Organization, Prospect Nauki 5, 142290 Pushchino, Moscow Region, Russian Federation.
Liubov
L
Trubitsina
G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Federal Agency of Scientific Organization, Prospect Nauki 5, 142290 Pushchino, Moscow Region, Russian Federation.
Alexander
A
Lisov
G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Federal Agency of Scientific Organization, Prospect Nauki 5, 142290 Pushchino, Moscow Region, Russian Federation.
Marina
M
Zakharova
G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Federal Agency of Scientific Organization, Prospect Nauki 5, 142290 Pushchino, Moscow Region, Russian Federation.
Alexey
A
Leontievsky
D000595:Amino Acid Sequence; D003460:Crystallization; D042845:Laccase / Q000737:chemistry*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D013056:Spectrophotometry, Ultraviolet; D055672:Static Electricity; D013302:Streptomyces / Q000201:enzymology*; D014961:X-Ray Diffraction
D042845:Laccase
Streptomyces griseoflavus; two-domain laccase