A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox
Contribuinte(s) |
UNIVERSIDADE DE SÃO PAULO |
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Data(s) |
17/04/2012
17/04/2012
2011
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Resumo |
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. FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo)[2005/54855-0] FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo)[2007/51700-1] FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo)[2009/10454-3] |
Identificador |
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, v.67, p.475-478, 2011 1744-3091 http://producao.usp.br/handle/BDPI/14883 10.1107/S1744309111003770 |
Idioma(s) |
eng |
Publicador |
WILEY-BLACKWELL |
Relação |
Acta Crystallographica Section F-structural Biology and Crystallization Communications |
Direitos |
closedAccess Copyright WILEY-BLACKWELL |
Palavras-Chave | #seeding #crystallization in the presence of oil #l-amino-acid oxidases #snake venoms #Bothrops atrox #VIPER CALLOSELASMA-RHODOSTOMA #HUMAN PLATELET-AGGREGATION #SNAKE-VENOM #PROTEIN CRYSTALLIZATION #PURIFICATION #ANTIBACTERIAL #MECHANISM #LAAO |
Tipo |
article original article publishedVersion |