title: A rational protocol for the successful crystallization of L-amino-acid oxidase from Bothrops atrox.
abstract: Despite the valuable contributions of robotics and high-throughput approaches to protein crystallization, the role of an experienced crystallographer in the evaluation and rationalization of a crystallization process is still crucial to obtaining crystals suitable for X-ray diffraction measurements. In this work, the difficult task of crystallizing the flavoenzyme L-amino-acid oxidase purified from Bothrops atrox snake venom was overcome by the development of a protocol that first required the identification of a non-amorphous precipitate as a promising crystallization condition followed by the implementation of a methodology that combined crystallization in the presence of oil and seeding techniques. Crystals were obtained and a complete data set was collected to 2.3 Å resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.64, b = 123.92, c = 105.08 Å, β = 96.03°. There were four protein subunits in the asymmetric unit, which gave a Matthews coefficient V(M) of 2.12 Å(3) Da(-1), corresponding to 42% solvent content. The structure has been solved by molecular-replacement techniques.
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirao Preto-FCFRP-USP, Avenida do Café s/n, Ribeirão Preto, 14040-903 São Paulo, Brazil.
authors.forename: Raquel Melo
authors.initials: RM
authors.lastname: Alves
authors.forename: Patricia Rosa
authors.initials: PR
authors.lastname: Feliciano
authors.forename: Suely Vilela
authors.initials: SV
authors.lastname: Sampaio
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Nonato
mesh_terms: D000818:Animals; D017837:Bothrops; D003460:Crystallization; D018360:Crystallography, X-Ray; D050830:L-Amino Acid Oxidase / Q000737:chemistry*
chemical_list: D050830:L-Amino Acid Oxidase