title: Crystallization and crystallographic analysis of branching enzymes from Cyanothece sp. ATCC 51142.
abstract: Several cyanobacterial species, including Cyanothece sp. ATCC 51142, remarkably have four isoforms of α-glucan branching enzymes (BEs). Based on their primary structures, they are classified into glycoside hydrolase (GH) family 13 (BE1, BE2 and BE3) or family 57 (GH57 BE). In the present study, GH13-type BEs from Cyanothece sp. ATCC 51142 (BE1, BE2 and BE3) have been overexpressed in Escherichia coli and biochemically characterized. The recombinant BE1 was crystallized by the hanging-drop vapour-diffusion method. Crystals of BE1 were obtained at 293 K in the presence of 0.2 M Mg(2+), 7-10%(w/v) ethanol, 0.1 M HEPES-NaOH pH 7.2-7.9. The crystals belonged to the tetragonal space group P41212, with unit-cell parameters a = b = 133.75, c = 185.90 Å, and diffracted to beyond 1.85 Å resolution. Matthews coefficient calculations suggested that the crystals of BE1 contained two molecules in the asymmetric unit.
authors.affiliation: Department of Biological Production, Faculty of Bioresource Sciences, Akita Prefectural University, Shimoshinjyo-Nakano, Akita 010-0195, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Hayashi
authors.affiliation: Department of Biological Production, Faculty of Bioresource Sciences, Akita Prefectural University, Shimoshinjyo-Nakano, Akita 010-0195, Japan.
authors.forename: Ryuichiro
authors.initials: R
authors.lastname: Suzuki
authors.affiliation: Unité de Glycobiologie Structurale et Fonctionnelle, Unité Mixte de Recherche 8576, Centre National de la Recherche Scientifique-Université des Sciences et Technologies de Lille, 59655 Villeneuve d'Ascq CEDEX, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Colleoni
authors.affiliation: Unité de Glycobiologie Structurale et Fonctionnelle, Unité Mixte de Recherche 8576, Centre National de la Recherche Scientifique-Université des Sciences et Technologies de Lille, 59655 Villeneuve d'Ascq CEDEX, France.
authors.forename: Steven G
authors.initials: SG
authors.lastname: Ball
authors.affiliation: Department of Biological Production, Faculty of Bioresource Sciences, Akita Prefectural University, Shimoshinjyo-Nakano, Akita 010-0195, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Fujita
authors.affiliation: Department of Biological Production, Faculty of Bioresource Sciences, Akita Prefectural University, Shimoshinjyo-Nakano, Akita 010-0195, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Suzuki
mesh_terms: D015061:1,4-alpha-Glucan Branching Enzyme / Q000737:chemistry* / Q000235:genetics; D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D046889:Cyanothece / Q000737:chemistry* / Q000201:enzymology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D010957:Plasmids; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D015061:1,4-alpha-Glucan Branching Enzyme
keywords: Cyanothece; amylopectin; branching enzyme; cyanobacteria; glycogen