Crystallization and preliminary X-ray analysis of a novel Kunitz-type kallikrein inhibitor from Bauhinia bauhinioides

Crystallization and preliminary X-ray analysis of a novel Kunitz-type kallikrein inhibitor from Bauhinia bauhinioides

Autor Navarro, MVDS Google Scholar
Vierira, D. F. Google Scholar
Nagem, RAP Google Scholar
Araujo, APU de Google Scholar
Oliva, MLV Google Scholar
Garratt, R. C. Google Scholar
Instituição Universidade de São Paulo (USP)
Universidade Federal de Minas Gerais (UFMG)
Universidade Federal de São Paulo (UNIFESP)
Resumo A Kunitz-type protease inhibitor (BbKI) found in Bauhinia bauhinioides seeds has been overexpressed in Escherichia coli and crystallized at 293 K using PEG 4000 as the precipitant. X-ray diffraction data have been collected to 1.87 A resolution using an in-house X-ray generator. the crystals of the recombinant protein (rBbKI) belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 46.70, b = 64.14, c = 59.24 angstrom. Calculation of the Matthews coefficient suggests the presence of one monomer of rBbKI in the asymmetric unit, with a corresponding solvent content of 51% (V(M) = 2.5 angstrom(3) Da(-1)). Iodinated crystals were prepared and a derivative data set was also collected at 2.1 angstrom resolution. Crystals soaked for a few seconds in a cryogenic solution containing 0.5 M NaI were found to be reasonably isomorphous to the native crystals. Furthermore, the presence of iodide anions could be confirmed in the NaI-derivatized crystal. Data sets from native and derivative crystals are being evaluated for use in crystal structure determination by means of the SIRAS (single isomorphous replacement with anomalous scattering) method.
Idioma Inglês
Data de publicação 2005-10-01
Publicado em Acta Crystallographica Section F-structural Biology and Crystallization Communications. Malden: Wiley-Blackwell, v. 61, p. 910-913, 2005.
ISSN 1744-3091 (Sherpa/Romeo, fator de impacto)
Publicador Wiley-Blackwell
Extensão 910-913
Fonte http://dx.doi.org/10.1107/S1744309105028496
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000232354300012
Endereço permanente http://repositorio.unifesp.br/handle/11600/28500

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