Crystallization and preliminary X-ray crystallographic studies of the D2 region of the skeletal muscle ryanodine receptor

Abstract

The N-terminal portion (amino acids 1303-1367) of the type 1 ryanodine receptor D2 region is thought to be critical for excitation-contraction coupling in skeletal muscle. A segment of the D2 region (amino acids 1317-1355) was expressed as a glutathione S-transferase fusion protein (GST-D2) and then crystallized at room temperature using ammonium sulfate as precipitant. Using a newly developed cryo-soaking method, complete native data sets were measured to a resolution of 2.2 Angstrom using synchrotron radiation. The crystal was found to be hexagonal, belonging to space group P6(3)22, with unit-cell parameters a = b = 116.1, c = 77.9 Angstrom.