Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex

dc.creatorTrejo, Sebastián Alejandro
dc.creatorLópez, Laura María Isabel
dc.creatorCaffini, Néstor Oscar
dc.creatorNatalucci, Claudia Luisa
dc.creatorCanals, Francesc
dc.creatorAvilés, Francesc X.
dc.date2009-07
dc.date2022-09-20T12:33:18Z
dc.date.accessioned2023-07-15T05:11:33Z
dc.date.available2023-07-15T05:11:33Z
dc.identifierhttp://sedici.unlp.edu.ar/handle/10915/142367
dc.identifierissn:1432-2048
dc.identifierissn:0032-0935
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/7472429
dc.descriptionAsclepain f is a papain-like protease previously isolated and characterized from latex of <i>Asclepias fruticosa</i>. This enzyme is a member of the C1 family of cysteine proteases that are synthesized as preproenzymes. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was determined by molecular modeling. A full-length 1,152 bp cDNA was cloned by RT-RACE-PCR from latex mRNA. The sequence was predicted as an open reading frame of 340 amino acid residues, of which 16 residues belong to the signal peptide, 113 to the propeptide and 211 to the mature enzyme. The full-length cDNA was ligated to pPICZα vector and expressed in <i>Pichia pastoris</i>. Recombinant asclepain f showed endopeptidase activity on pGlu-Phe-Leu-p-nitroanilide and was identified by PMF-MALDI-TOF MS. Asclepain f is the first peptidase cloned and expressed from mRNA isolated from plant latex, confirming the presence of the preprocysteine peptidase in the latex.
dc.descriptionCentro de Investigación de Proteínas Vegetales
dc.formatapplication/pdf
dc.format319-328
dc.languageen
dc.rightshttp://creativecommons.org/licenses/by/4.0/
dc.rightsCreative Commons Attribution 4.0 International (CC BY 4.0)
dc.subjectCiencias Exactas
dc.subjectBiología
dc.subjectAsclepias fruticosa
dc.subjectGomphocarpus fruticosus subsp. fruticosus
dc.subjectPlant latex
dc.subjectCysteine endopeptidase
dc.subjectCloning
dc.subjectOverexpression in Pichia pastoris
dc.titleSequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <i>Asclepias fruticosa</i> latex
dc.typeArticulo
dc.typeArticulo


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