Overexpression, crystallization and preliminary X-ray crystallographic analysis of the C-terminal cytosolic domain of mouse anoctamin 1
- Authors
- Park, Sang Ho; Chung, Ho Kyung; Kim, Do Jin; Han, Mi Ra; Park, Mi Seul; Oh, Uhtaek; Kim, Hyun-Jung; Han, Byung Woo
- Issue Date
- Oct-2011
- Publisher
- WILEY-BLACKWELL
- Keywords
- anoctamin 1; chloride channels; transmembrane protein 16A
- Citation
- ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, v.67, no.10, pp 1250 - 1252
- Pages
- 3
- Journal Title
- ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS
- Volume
- 67
- Number
- 10
- Start Page
- 1250
- End Page
- 1252
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/21237
- DOI
- 10.1107/S1744309111027989
- ISSN
- 1744-3091
2053-230X
- Abstract
- Transmembrane protein 16A (TMEM16A, also known as anoctamin 1; ANO1) is a bona fide Ca2+-activated chloride channel that is activated by intracellular Ca2+- and Ca2+-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 angstrom 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 angstrom. 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 angstrom(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.
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Collections - College of Pharmacy > School of Pharmacy > 1. Journal Articles
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