Journal of Protein Chemistry

, Volume 20, Issue 8, pp 625–632 | Cite as

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

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 mollis Mimosoideae trypsin inhibitor N-terminal sequence Kunitz family 

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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

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