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Funastrain c II: A Cysteine Endopeptidase Purified from the Latex of Funastrum clausum

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Abstract

A cysteine endopeptidase, named funastrain c II, was isolated and characterized from the latex of Funastrum clausum (Asclepiadaceae). The molecular mass (mass spectrometry) of the protease was 23.636 kDa. The analysis of funastrain c II by SDS-PAGE revealed a single polypeptide chain. The enzyme showed a remarkable stability of its caseinolytic activity after incubation at temperatures as high as 70°C. Inhibition and activation assays indicated the cysteinic nature of the funastrain c II catalytic site. The optimum pH of funastrain c II enzymatic activity varied according to the substrate used (9.0–10.0 for casein and 6.2–6.8 for PFLNA). Kinetic parameters were determined for N-α-CBZ-Ala p-nitrophenyl ester (K m = 0.0243 mM, k cat = 1.5 s−1) and l-pyroglutamyl-l-phenylalanyl-l-leucine-p-nitroanilide (PFLNA; K M = 0.1011 mM, k cat = 0.9 s−1). The N-terminal sequence of funastrain c II showed considerable similarity to other proteases isolated from latex of different Asclepiadaceae species as well as to other cysteine proteinases belonging to the papain family.

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Authors
  1. Susana R. Morcelle
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  2. Sebastián A. Trejo
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  3. Francesc Canals
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  4. Francesc X. Avilés
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  5. Nora S. Priolo
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Morcelle, S.R., Trejo, S.A., Canals, F. et al. Funastrain c II: A Cysteine Endopeptidase Purified from the Latex of Funastrum clausum . J Protein Chem 23, 205–215 (2004). https://doi.org/10.1023/B:JOPC.0000026416.90134.7b

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  • DOI: https://doi.org/10.1023/B:JOPC.0000026416.90134.7b

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