title: Human cardiac ryanodine receptor: preparation, crystallization and preliminary X-ray ANALysis of the N-terminal region.
abstract: Human ryanodine receptor 2 (hRyR2) is a calcium ion channel present in the membrane of the sarcoplasmic reticulum of cardiac myocytes that mediates release of calcium ions from the sarcoplasmic reticulum stores during excitation- contraction coupling. Disease-causing mutations of hRyR2 are clustered into N-terminal (amino acids 1-600), central (amino acids 2100-2500) and C-terminal (amino acids 3900-5000) regions. These regions are believed to be involved in regulation of channel gating. The N-terminal region of hRyR2 has been implicated in regulating basal channel activity by interaction with the central hRyR2 region. This paper reports preparation, crystallization and preliminary X-ray analysis of recombinant hRyR2(1-606) N-terminal fragment. Soluble hRyR2(1-606) was expressed in Escherichia coli. Purification conditions were optimized using thermal shift assay. The quality and stability of the sample was probed by dynamic light scattering. A monomeric protein showing over 95% purity was obtained. The protein was crystallized by the hanging drop vapor-diffusion method. Diffraction data with resolution 2.39 Å were collected and processed.
authors.affiliation: Institute of Molecular Biology SAS, Dúbravska cesta 21, 845 51 Bratislava. jozef.sevcik@savba.sk.
authors.forename: L'ubomír
authors.initials: L
authors.lastname: Borko
authors.forename: Július
authors.initials: J
authors.lastname: Košt'an
authors.forename: Alexandra
authors.initials: A
authors.lastname: Zahradníková
authors.forename: Vladimír
authors.initials: V
authors.lastname: Pevala
authors.forename: Juraj
authors.initials: J
authors.lastname: Gašperík
authors.forename: Eva
authors.initials: E
authors.lastname: Hostinová
authors.forename: L'ubica
authors.initials: L
authors.lastname: Urbániková
authors.forename: Kristina
authors.initials: K
authors.lastname: Djinović-Carugo
authors.forename: Vladena
authors.initials: V
authors.lastname: Bauerová-Hlinková
authors.forename: Jozef
authors.initials: J
authors.lastname: Sevčík
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D006801:Humans; D018482:Muscle, Skeletal / Q000737:chemistry*; D009206:Myocardium / Q000737:chemistry; D032383:Myocytes, Cardiac / Q000737:chemistry*; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D012519:Sarcoplasmic Reticulum / Q000737:chemistry
chemical_list: D019837:Ryanodine Receptor Calcium Release Channel