A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox

作者：Alves Raquel Melo; Feliciano Patricia Rosa; Sampaio Suely Vilela; Nonato Maria Cristina^*

来源：Acta Crystallographica Section F-Structural Biology and Crystallization Communications, 2011, 67: 475-478.

DOI：10.1107/S1744309111003770

• seeding
• crystallization in the presence of oil
• l-amino-acid oxidases
• snake venoms
• Bothrops atrox

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 A resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.64, b = 123.92, c = 105.08 A, beta = 96.03 degrees. There were four protein subunits in the asymmetric unit, which gave a Matthews coefficient V (M) of 2.12 A3 Da-1, corresponding to 42% solvent content. The structure has been solved by molecular-replacement techniques.

• 出版日期2011-4