Crystallization and preliminary X-ray diffraction studies on the bicupin YwfC from Bacillus subtilis

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2006-12, Vol.62 (12), p.1259-1262
• Main Authors: Rajavel, M., Gopal, B.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Bacillus subtilis; Bacillus subtilis - chemistry; Bacterial Proteins - chemistry; CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTAL STRUCTURE; CRYSTALLIZATION; Crystallization Communications; Crystallography, X-Ray; CRYSTALS; cupin fold; DIAMONDS; EVOLUTION; functional diversity; gene duplication; Molecular Sequence Data; MOLECULES; OXALATES; OXYGEN; Protein Structure, Tertiary; RESOLUTION; Sequence Alignment; SOLVENTS; SPACE GROUPS; VARIATIONS; X-RAY DIFFRACTION
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309106047087

A central tenet of evolutionary biology is that proteins with diverse biochemical functions evolved from a single ancestral protein. A variation on this theme is that the functional repertoire of proteins in a living organism is enhanced by the evolution of single‐chain multidomain polypeptides by gene‐fusion or gene‐duplication events. Proteins with a double‐stranded β‐helix (cupin) scaffold perform a diverse range of functions. Bicupins are proteins with two cupin domains. There are four bicupins in Bacillus subtilis, encoded by the genes yvrK, yoaN, yxaG and ywfC. The extensive phylogenetic information on these four proteins makes them a good model system to study the evolution of function. The proteins YvrK and YoaN are oxalate decarboxylases, whereas YxaG is a quercetin dioxygenase. In an effort to aid the functional annotation of YwfC as well as to obtain a complete structure–function data set of bicupins, it was proposed to determine the crystal structure of YwfC. The bicupin YwfC was crystallized in two crystal forms. Preliminary crystallographic studies were performed on the diamond‐shaped crystals, which belonged to the tetragonal space group P422. These crystals were grown using the microbatch method at 298 K. Native X‐ray diffraction data from these crystals were collected to 2.2 Å resolution on a home source. These crystals have unit‐cell parameters a = b = 68.7, c = 211.5 Å. Assuming the presence of two molecules per asymmetric unit, the VM value was 2.3 Å3 Da−1 and the solvent content was approximately 45%. Although the crystals appeared less frequently than the tetragonal form, YwfC also crystallizes in the monoclinic space group P21, with unit‐cell parameters a = 46.7, b = 106.3, c = 48.7 Å, β = 92.7°.