Crystallization and preliminary X-ray crystallographic analysis of Ca{sup 2+}-independent and Ca{sup 2+}-dependent species of the type II antifreeze protein

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 62; no. Pt 6
• Main Authors: Nishimiya, Yoshiyuki, Kondo, Hidemasa, Yasui, Masanori, Division of Biological Sciences, Graduate School of Science, Hokkaido University, N8W5, Kita, Sapporo 060-0808, Sugimoto, Hiroshi, Noro, Natsuko, Sato, Ryoko, Suzuki, Mamoru, Miura, Ai, Tsuda, Sakae
• Format: Journal Article
• Language: English
• Published: United Kingdom 01.06.2006
• Subjects: BOUND STATE; CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; DIFFRACTION; DIFFUSION; PROTEINS; RESOLUTION; SPACE GROUPS; SYMMETRY
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309106015570

Crystallization and X-ray data analyses were successful for both Ca{sup 2+}-independent and Ca{sup 2+}-dependent species of the type II antifreeze protein. The resolution of the crystal was 1.35 Å for the Ca{sup 2+}-independent species, and was 1.25 and 1.06 Å for the Ca{sup 2+}-dependent species in the Ca{sup 2+}-free and -bound states, respectively. Ca{sup 2+}-independent and Ca{sup 2+}-dependent species of the type II antifreeze protein (AFP) were both crystallized using the hanging-drop vapour-diffusion method. It appeared that the crystal of the Ca{sup 2+}-independent species from Brachyosis rostratus belongs to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 43.3, b = 48.4, c = 59.7 Å, and diffraction data were collected to 1.34 Å resolution. For the Ca{sup 2+}-dependent type II AFP species from Hypomesus nipponensis, crystallization was carried out for its Ca{sup 2+}-free and Ca{sup 2+}-bound states. 1.25 Å resolution data were collected from the crystal in the Ca{sup 2+}-free state, which exhibited P3{sub 1}21 (or P3{sub 2}21) symmetry, with unit-cell parameters a = b = 66.0, c = 50.3 Å. Data collection could be extended to 1.06 Å resolution for the crystal in the Ca{sup 2+} -bound state, which appeared to be isomorphous to the crystal in the Ca{sup 2+}-free state (unit-cell parameters a = b = 66.0, c = 49.8 Å). These data will allow us to determine the high-resolution structures of the two species of type II AFP.