title: Purification, crystallization and preliminary crystallographic analysis of the full-length cystathionine β-synthase from Apis mellifera.
abstract: Cystathionine β-synthase (CBS) is a pyridoxal-5'-phosphate-dependent enzyme that catalyzes the first step of the transsulfuration pathway, namely the condensation of serine with homocysteine to form cystathionine. Mutations in the CBS gene are the single most common cause of hereditary homocystinuria, a multisystemic disease affecting to various extents the vasculature, connective tissues and central nervous system. At present, the crystal structure of CBS from Drosophila melanogaster is the only available structure of the full-length enzyme. Here we describe a cloning, overexpression, purification and preliminary crystallographic analysis of a full-length CBS from Apis mellifera (AmCBS) which maintains 51 and 46% sequence identity with its Drosophila and human homologs, respectively. The AmCBS yielded crystals belonging to space group P2(1)2(1)2(1), with unit-cell parameters a=85.90, b=95.87, c=180.33 Å. Diffraction data were collected to a resolution of 3.0 Å. The crystal structure contained two molecules in the asymmetric unit which presumably correspond to the dimeric species observed in solution.
authors.affiliation: Structural Biology Unit, CIC bioGUNE, Parque Tecnológico de Bizkaia, Edificio 800, Derio, Bizkaia 48160, Spain.
authors.forename: Iker
authors.initials: I
authors.lastname: Oyenarte
authors.forename: Tomas
authors.initials: T
authors.lastname: Majtan
authors.forename: June
authors.initials: J
authors.lastname: Ereño
authors.forename: María Angeles
authors.initials: MA
authors.lastname: Corral-Rodríguez
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Klaudiny
authors.forename: Juraj
authors.initials: J
authors.lastname: Majtan
authors.forename: Jan P
authors.initials: JP
authors.lastname: Kraus
authors.forename: Luis Alfonso
authors.initials: LA
authors.lastname: Martínez-Cruz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001516:Bees / Q000201:enzymology*; D017124:Conserved Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D003541:Cystathionine beta-Synthase / Q000737:chemistry* / Q000302:isolation & purification; D019476:Insect Proteins / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
chemical_list: D019476:Insect Proteins; D003541:Cystathionine beta-Synthase