Cloning, purification, crystallization and preliminary X-ray diffraction analysis of the antistasin-type inhibitor ghilanten (domain I) from Haementeria ghilianii in complex with porcine beta-trypsin

Cloning, purification, crystallization and preliminary X-ray diffraction analysis of the antistasin-type inhibitor ghilanten (domain I) from Haementeria ghilianii in complex with porcine beta-trypsin

Autor Rester, U. Google Scholar
Bode, W. Google Scholar
Sampaio, CAM Google Scholar
Auerswald, E. A. Google Scholar
Lopes, APY Google Scholar
Instituição Max Planck Inst Biochem
Universidade Federal de São Paulo (UNIFESP)
Univ Munich
Univ Munich Klinikum
Inst Butantan
Resumo Ghilanten, isolated from the leech Haementeria ghilianii, is a potent two-domain anticoagulant protein homologous to the factor Xa inhibitor antistasin. A synthetic gene encoding the amino-terminal domain of ghilanten (ghilanten-D1) was constructed, expressed in the methylotrophic yeast Pichia pastoris and purified by heparin-Sepharose chromatography. Recombinant ghilanten-D1 inhibits bovine trypsin and human factor Xa with equilibrium inhibition constants (K-i) of 126 and 1.2 nM, respectively. Ghilanten-D1 has been crystallized in complex with porcine beta -trypsin; three different-looking but isomorphous crystal forms were obtained, each belonging to the orthorhombic space group P2(1)2(1)2(1). These crystals diffracted to beyond 3.6 Angstrom resolution using a rotating-anode X-ray source. A data set complete to 3.7 Angstrom resolution was collected.
Idioma Inglês
Data de publicação 2001-07-01
Publicado em Acta Crystallographica Section D-biological Crystallography. Copenhagen: Munksgaard Int Publ Ltd, v. 57, p. 1038-1041, 2001.
ISSN 0907-4449 (Sherpa/Romeo, fator de impacto)
Publicador Munksgaard Int Publ Ltd
Extensão 1038-1041
Fonte http://dx.doi.org/10.1107/S0907444901007272
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000169534700017
Endereço permanente http://repositorio.unifesp.br/handle/11600/26592

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