title: Crystallization of the coiled-coil domain of Atg16 essential for autophagy.
abstract: Atg16 is a scaffold protein that interacts with Atg12-Atg5 protein conjugates via its N-terminal domain and self-assembles via its coiled-coil domain, thus forming a multimeric Atg12-Atg5-Atg16 complex that is essential for autophagy. The coiled-coil domain of Atg16 was expressed, purified and crystallized. The crystal belonged to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = 127.7, c = 77.8 A. Self-rotation functions and volume-to-weight ratio values suggested that the crystal may contain six molecules per asymmetric unit. Since the domain does not contain a methionine residue, selenomethionine-labelled crystals were prepared with a leucine-to-methionine substitution in the coiled-coil domain and these crystals were used for the collection of single-wavelength anomalous dispersion data to 2.5 A resolution.
authors.affiliation: Department of Structural Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, N-21, W-11, Kita-ku, Sapporo 001-0021, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Fujioka
authors.forename: Nobuo N
authors.initials: NN
authors.lastname: Noda
authors.forename: Minako
authors.initials: M
authors.lastname: Matsushita
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ohsumi
authors.forename: Fuyuhiko
authors.initials: F
authors.lastname: Inagaki
mesh_terms: D000595:Amino Acid Sequence; D001343:Autophagy; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins / Q000737:chemistry*; D003460:Crystallization; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000737:chemistry; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: C498649:ATG16 protein, S cerevisiae; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins; D046912:Multiprotein Complexes; D029701:Saccharomyces cerevisiae Proteins