Open AccessPurification, crystallization and preliminary crystallographic analysis of the full‐length cystathionine β‐synthase from Apis mellifera
Author(s) -
Oyenarte Iker,
Majtan Tomas,
Ereño June,
CorralRodríguez María Angeles,
Klaudiny Jaroslav,
Majtan Juraj,
Kraus Jan P.,
MartínezCruz Luis Alfonso
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112038638
Subject(s) - cystathionine beta synthase , homocystinuria , crystallography , pyridoxal phosphate , space group , transsulfuration , chemistry , crystallization , crystal structure , lyase , molecule , biology , stereochemistry , enzyme , biochemistry , methionine , x ray crystallography , amino acid , diffraction , cofactor , physics , organic chemistry , optics
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 ( Am CBS) which maintains 51 and 46% sequence identity with its Drosophila and human homologs, respectively. The Am CBS yielded crystals belonging to space group P 2 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.