Print Email Facebook Twitter Structural characterization of potato protease inhibitor I (cv. Bintje) after expression in Pichia pastoris Title Structural characterization of potato protease inhibitor I (cv. Bintje) after expression in Pichia pastoris Author TNO Zeist van den Broek, L.A.M. Pouvreau, L. Lommerse, G. Schipper, B. van Koningsveld, G.A. Gruppen, H. Publication year 2004 Abstract In the present study the structural properties of potato protease inhibitor 1 (PI-1) were studied as a function of temperature to elucidate its precipitation mechanism upon heating. A cDNA coding for PI-1 from cv. Bintje was cloned and expressed in Pichia pastoris. Using the recombinant PI-1 it was suggested that PI-1 behaves as a hexameric protein rather than as a pentamer, as previously proposed in the literature. The recombinant protein seems either to have a predominantly unordered structure or to belong to the β-II proteins. Differential scanning calorimetry analysis of PI-1 revealed that its thermal unfolding occurs via one endothermic transition in which the hexameric PI-1 probably unfolds, having a dimer instead of a monomer as cooperative unit. The transition temperature for the recombinant PI-1 was 88 °C. Similar results were obtained for a partially purified pool of native PI-1 from cv. Bintje. Molecular Sequence Numbers: GENBANK: AY496262; Chemicals / CAS: proteinase inhibitor, 37205-61-1; Plant Proteins; Protease Inhibitors; Recombinant Proteins Subject PotatoProtease inhibitorRecombinant PI-1Solanum tuberosumcomplementary DNAproteinase inhibitor 1recombinant proteinunclassified drugdifferential scanning calorimetryenzyme inhibitiongene expressionnonhumannucleotide sequencePichia pastorispolyacrylamide gel electrophoresispolymerase chain reactionprotein purificationproteinase inhibitionroot vegetableCalorimetry, Differential ScanningChemistry, PhysicalCircular DichroismDrug StabilityGene ExpressionHeatMolecular WeightPichiaPlant ProteinsProtease InhibitorsProtein FoldingRecombinant ProteinsSolanum tuberosumPichiaPichia pastorisSolanumSolanum tuberosum To reference this document use: http://resolver.tudelft.nl/uuid:51311c2b-5ba1-4258-a616-5c287d4da51c DOI https://doi.org/10.1021/jf049832x TNO identifier 237906 ISSN 0021-8561 Source Journal of Agricultural and Food Chemistry, 52 (52), 4928-4934 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.