Journal of Protein Chemistry

, Volume 20, Issue 8, pp 625–632

Purification and Characterization of a New Trypsin Inhibitor from Dimorphandra mollis Seeds

  • Gláucia C. Mello
  • Maria Luiza V. Oliva
  • Joana T. Sumikawa
  • Olga L. T. Machado
  • Sérgio Marangoni
  • José C. Novello
  • Maria Lígia R. Macedo
Article

DOI: 10.1023/A:1013764118579

Cite this article as:
Mello, G.C., Oliva, M.L.V., Sumikawa, J.T. et al. J Protein Chem (2001) 20: 625. doi:10.1023/A:1013764118579

Abstract

A second trypsin inhibitor (DMTI-II) was purified from the seed of Dimorphandra mollis (Leguminosae-Mimosoideae) by ammonium sulfate precipitation (30–60%), gel filtration, and ion-exchange and affinity chromatography. A molecular weight of 23 kDa was estimated by gel filtration on a Superdex 75 column SDS-PAGE under reduced conditions showed that DMTI-II consisted of a single polypeptide chain, although isoelectric focusing revealed the presence of three isoforms. The dissociation constant of 1.7 × 10−9 M with bovine trypsin indicated a high affinity between the inhibitor and this enzyme. The inhibitory activity was stable over a wide pH range and in the presence of DTT. The N-terminal sequence of DMTI-II showed a high degree of homology with other Kunitz-type inhibitors.

Dimorphandra mollisMimosoideaetrypsin inhibitorN-terminal sequenceKunitz family

Copyright information

© Plenum Publishing Corporation 2001

Authors and Affiliations

  • Gláucia C. Mello
    • 1
  • Maria Luiza V. Oliva
    • 1
  • Joana T. Sumikawa
    • 1
  • Olga L. T. Machado
    • 2
  • Sérgio Marangoni
    • 3
  • José C. Novello
    • 3
  • Maria Lígia R. Macedo
    • 3
  1. 1.Departamento de Bioquímica, Instituto de BiologiaUniversidade Estadual de Campinas (UNICAMP)Campinas, SPBrazil
  2. 2.Centro de Biociências e BiotecnologiaUniversidade Estadual do Norte FluminenseCampo dos Goytacazes, RJBrazil
  3. 3.Departamento de Bioquímica, Instituto de BiologiaUniversidade Estadual de Campinas (UNICAMP)Campinas, SPBrazil
  4. 4.Departamento de Ciências NaturaisUniversidade Federal de Mato Grosso do Sul (CEUL/UFMS)Três Lagoas, MSBrazil