Purification and Characterization of a New Plant Endopeptidase Isolated from Latex of Asclepias fruticosa L. (Asclepiadaceae)

Asclepias fruticosa L. is a small shrub containing latex with proteolytic activity. The crude extract (latex diluted 1:250 and ultracentrifuged) contained 276 μg of protein/mL and the proteolytic activity reached 1.2 caseinolytic U/mL. This enzyme preparation was very stable even after 2 hours at 45...

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Detalles Bibliográficos
Autores principales: Trejo, Sebastián Alejandro, López, Laura María Isabel, Cimino, Cecilia Verónica, Caffini, Néstor Oscar, Natalucci, Claudia Luisa
Formato: Articulo
Lenguaje:Inglés
Publicado: 2001
Materias:
Biología
Asclepias fruticosa
Asclepiadaceae
Latex
Milkweed
Plant endopeptidases
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/132505
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-132505
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Biología
Asclepias fruticosa
Asclepiadaceae
Latex
Milkweed
Plant endopeptidases
spellingShingle Biología
Asclepias fruticosa
Asclepiadaceae
Latex
Milkweed
Plant endopeptidases
Trejo, Sebastián Alejandro
López, Laura María Isabel
Cimino, Cecilia Verónica
Caffini, Néstor Oscar
Natalucci, Claudia Luisa
Purification and Characterization of a New Plant Endopeptidase Isolated from Latex of Asclepias fruticosa L. (Asclepiadaceae)
topic_facet Biología
Asclepias fruticosa
Asclepiadaceae
Latex
Milkweed
Plant endopeptidases
description Asclepias fruticosa L. is a small shrub containing latex with proteolytic activity. The crude extract (latex diluted 1:250 and ultracentrifuged) contained 276 μg of protein/mL and the proteolytic activity reached 1.2 caseinolytic U/mL. This enzyme preparation was very stable even after 2 hours at 45°C, but was quickly inactivated after 5 minutes at 80°C. Chromatographic purification was achieved by FPLC using a cation exchanger (SP-Sepharose FF). Thus, a unique proteolitically active fraction could be isolated, being homogeneous by bidimensional electrophoresis and mass spectrometry (Mr = 23,652). The optimum pH range was achieved at 8.5–10.5. The enzyme activity was completely inhibited by specific cysteine peptidases inhibitors. Isoelectric focusing followed by zymogram showed the enzyme had a pI greater than 9.3. The N-terminus sequence (LPDSVDWREKGVVFPIRNQGK) shows a great deal of similarity to those of the other cysteine endopeptidases isolated from latices of Asclepiadaceae even when a high degree of homology could be observed with other plant cysteine endopeptidases.
format Articulo
Articulo
author Trejo, Sebastián Alejandro
López, Laura María Isabel
Cimino, Cecilia Verónica
Caffini, Néstor Oscar
Natalucci, Claudia Luisa
author_facet Trejo, Sebastián Alejandro
López, Laura María Isabel
Cimino, Cecilia Verónica
Caffini, Néstor Oscar
Natalucci, Claudia Luisa
author_sort Trejo, Sebastián Alejandro
title Purification and Characterization of a New Plant Endopeptidase Isolated from Latex of Asclepias fruticosa L. (Asclepiadaceae)
title_short Purification and Characterization of a New Plant Endopeptidase Isolated from Latex of Asclepias fruticosa L. (Asclepiadaceae)
title_full Purification and Characterization of a New Plant Endopeptidase Isolated from Latex of Asclepias fruticosa L. (Asclepiadaceae)
title_fullStr Purification and Characterization of a New Plant Endopeptidase Isolated from Latex of Asclepias fruticosa L. (Asclepiadaceae)
title_full_unstemmed Purification and Characterization of a New Plant Endopeptidase Isolated from Latex of Asclepias fruticosa L. (Asclepiadaceae)
title_sort purification and characterization of a new plant endopeptidase isolated from latex of asclepias fruticosa l. (asclepiadaceae)
publishDate 2001
url http://sedici.unlp.edu.ar/handle/10915/132505
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