Structural characterization of potato protease inhibitor I (Cv. Bintje) after expression in Pichia pastoris.
| Autor: | van den Broek LA; Centre for Protein Technology TNO-WU, P.O. Box 8129, 6700 EV Wageningen, The Netherlands., Pouvreau L, Lommerse G, Schipper B, Van Koningsveld GA, Gruppen H |
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| Jazyk: | angličtina |
| Zdroj: | Journal of agricultural and food chemistry [J Agric Food Chem] 2004 Jul 28; Vol. 52 (15), pp. 4928-34. |
| DOI: | 10.1021/jf049832x |
| Abstrakt: | 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 beta-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 degrees C. Similar results were obtained for a partially purified pool of native PI-1 from cv. Bintje. (Copyright 2004 American Chemical Society) |
| Databáze: | MEDLINE |
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