title: Overexpression, crystallization and preliminary X-ray crystallographic analysis of the C-terminal cytosolic domain of mouse anoctamin 1.
abstract: Transmembrane protein 16A (TMEM16A, also known as anoctamin 1; ANO1) is a bona fide Ca(2+)-activated chloride channel that is activated by intracellular Ca(2+)- and Ca(2+)-mobilizing stimuli and plays important roles in a variety of physiological functions. To elucidate the structural features of ANO1, structural analysis of the C-terminal cytosolic domain of mouse ANO1 (mANO1-CTD) was initiated. mANO1-CTD was overexpressed in Escherichia coli and was crystallized at 297 K using a reservoir solution consisting of 0.2 M sodium acetate trihydrate, 0.1 M Tris-HCl pH 8.5 and 30%(w/v) PEG 4000. X-ray diffraction data were collected to 2.3 Å resolution. The crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 73.96, b = 103.73, c = 114.71 Å. If it is assumed that eight copies of a monomer molecule are present in the crystallographic asymmetric unit, the crystal volume per protein mass (V(M)) is 2.38 Å(3) Da(-1) and the solvent content is 48.38%. Attempts to solve the structure of mANO1-CTD by the MAD method using selenomethionine-labelled mANO1-CTD or heavy-atom-derivatized crystals are in progress.
authors.affiliation: Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Sang Ho
authors.initials: SH
authors.lastname: Park
authors.forename: Ho Kyung
authors.initials: HK
authors.lastname: Chung
authors.forename: Do Jin
authors.initials: DJ
authors.lastname: Kim
authors.forename: Mi Ra
authors.initials: MR
authors.lastname: Han
authors.forename: Mi Seul
authors.initials: MS
authors.lastname: Park
authors.forename: Uhtaek
authors.initials: U
authors.lastname: Oh
authors.forename: Hyun Jung
authors.initials: HJ
authors.lastname: Kim
authors.forename: Byung Woo
authors.initials: BW
authors.lastname: Han
mesh_terms: D000818:Animals; D000075369:Anoctamin-1; D018118:Chloride Channels / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D003600:Cytosol / Q000737:chemistry*; D015870:Gene Expression; D051379:Mice
chemical_list: C531181:ANO1 protein, mouse; D000075369:Anoctamin-1; D018118:Chloride Channels
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