A Kunitz-type glycosylated elastase inhibitor with one disulfide bridge

A Kunitz-type glycosylated elastase inhibitor with one disulfide bridge

Autor Sumikawa, J. T. Google Scholar
Nakahata, A. M. Google Scholar
Fritz, H. Google Scholar
Mentele, R. Google Scholar
Sampaio, M. U. Google Scholar
Oliva, MLV Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
LMU
Max Planck Inst Biochem
Resumo A glycosylated Bauhinia rufa elastase inhibitor (gBrEI) was purified and characterized using acetone precipitation, affinity chromatography on concanavalin A-Sepharose, ion-exchange chromatography on a HiTrap Q column, size exclusion chromatography on a Superdex 200 column and reverse-phase chromatography on a C-18 column. gBrEI inhibited pancreatic porcine elastase with an equilibrium dissociation constant (K-i) of 6.18 x 10(-8) M, but it did not inhibit human neutrophil elastase, bovine trypsin, human plasma kallikrein or porcine pancreatic kallikrein. On SDS-electrophoresis, gBrEI appeared as a single 20-kDa band, also after reduction. Schiff reagent staining indicated a carbohydrate portion in the protein, which was confirmed by mass spectrometry. the glycosylated site was Asn(38), and a carbohydrate portion of 1.17 kDa was identified. gBrEI was found to contain 144 amino acid residues, and a FASTA database analysis showed that it belongs to the plant Kunitz-type inhibitor family. Val(66) was identified as reactive site P1 residue by comparison of conserved positions in the sequences. Since gBrEI harbors a single disulfide bridge, it may be considered a new type of Kunitz inhibitor, intermediate between the classical kunitz inhibitors, which contain two disulfide bridges, and those from B. bauhinioides, which do not have such bridges.
Palavra-chave Bauhinia rufa
elastase/antagonists and inhibitors
glycoproteins
Kunitz inhibitor
leguminosae
plant
serine endopeptidases
Idioma Inglês
Data de publicação 2006-04-01
Publicado em Planta Medica. Stuttgart: Georg Thieme Verlag Kg, v. 72, n. 5, p. 393-397, 2006.
ISSN 0032-0943 (Sherpa/Romeo, fator de impacto)
Publicador Georg Thieme Verlag Kg
Extensão 393-397
Fonte http://dx.doi.org/10.1055/s-2005-916237
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000237192200002
Endereço permanente http://repositorio.unifesp.br/handle/11600/28827

Exibir registro completo




Arquivo

Arquivo Tamanho Formato Visualização

Não existem arquivos associados a este item.

Este item está nas seguintes coleções

Buscar


Navegar

Minha conta