Human Cardiac Ryanodine Receptor: Preparation, Crystallization and Preliminary X-ray Analysis of the N-terminal Region

• DOI: 10.2174/0929866511320110004
• Publication Date: 2013-09-18T12:53:12Z
• Authors: L’ubomír Borko; Július Kostan; Alexandra Zahradn...; Vladimír Pevala; Juraj Gasperík; Eva Hostinova; L’ubica Urbanikova; Kristina Djinovic...; Vladena Bauerova-...; Jozef Sevcík
• Keywords: Sarcoplasm

Human ryanodine receptor 2 (hRyR2) is a calcium ion channel present in the membrane of sarcoplasmic reticulum cardiac myocytes that mediates release ions from stores during excitation- contraction coupling. Disease-causing mutations hRyR2 are clustered into N-terminal (amino acids 1-600), central 2100-2500) and C-terminal 3900-5000) regions. These regions believed to be involved regulation gating. The region has been implicated regulating basal activity by interaction with region. This paper reports preparation, crystallization preliminary X-ray analysis recombinant hRyR2(1-606) fragment. Soluble was expressed Escherichia coli. Purification conditions were optimized using thermal shift assay. quality stability sample probed dynamic light scattering. A monomeric protein showing over 95% purity obtained. crystallized hanging drop vapor-diffusion method. Diffraction data resolution 2.39 Å collected processed.