Characterization of the proteolytic system present in Vasconcellea quercifolia latex

María José Torres, Sebastián Alejandro Trejo, Walter David Obregón, Francesc Xavier Avilés, Laura María Isabel López, Claudia Luisa Natalucci

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11 Citations (Scopus)

Abstract

Vasconcellea quercifolia (Caricaceae) latex contains several cysteine endopeptidases with high proteolytic activity. Cysteine endopeptidases are the main active compounds used by the plant as a defense mechanism. A proteolytic preparation from V. quercifolia ("oak leaved papaya") latex was purified by cation exchange chromatography. From SDS-PAGE and blotting of the selected fractions, the N-terminal amino acid sequences of polypeptides were determined by Edman's degradation. The analysis by peptide mass fingerprinting (PMF) of the enzymes allowed their characterization and confirmed the presence of seven different cysteine proteinases in the latex of V. quercifolia. Moreover, the comparison between the tryptic maps with those deposited in databases using the MASCOT tool showed that none of the isolated proteases matched with another plant protease. Notably, a propeptidase was detected in the plant latex, which is being the first report in this sense. Furthermore, the cDNA of one of the cysteine proteases that is expressed in the latex of V. quercifolia was cloned and sequenced. The consensus sequence was aligned using the ClustalX web server, which allowed detecting a high degree of identity with cysteine proteases of the Caricaceae family and establishing the evolutionary relationship between them. We also observed a high conservation degree for those amino acid residues which are essential for the catalytic activity and tridimensional structure of the plant proteases belonging to the subfamily C1A. The PMF analysis strongly suggests that the sequence obtained corresponds to the VQ-III peptidase. © 2012 Springer-Verlag.
Original languageEnglish
Pages (from-to)1471-1484
JournalPlanta
Volume236
Issue number5
DOIs
Publication statusPublished - 1 Oct 2012

Keywords

  • Caricaceae
  • cDNA
  • Cysteine endopeptidases
  • Peptide mass fingerprint
  • Plant proteases
  • Vasconcellea

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