Structural characterization of potato protease inhibitor I (Cv. Bintje) after expression in Pichia pastoris

L.A.M. van den Broek; L.A.M. Pouvreau; G. Lommerse; B. Schipper; G.A. van Koningsveld; H. Gruppen

doi:10.1021/jf049832x

Structural characterization of potato protease inhibitor I (Cv. Bintje) after expression in Pichia pastoris

L.A.M. van den Broek, L.A.M. Pouvreau, G. Lommerse, B. Schipper, G.A. van Koningsveld, H. Gruppen

Research output: Contribution to journal › Article › Academic › peer-review

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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 -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

Original language	English
Pages (from-to)	4928-4934
Journal	Journal of Agricultural and Food Chemistry
Volume	52
DOIs	https://doi.org/10.1021/jf049832x
Publication status	Published - 2004

Keywords

circular-dichroism
stability
proteins
chymotrypsin
tubers
solubility
elkana

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10.1021/jf049832x

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@article{377533e135034a05b7e43f674d6292fd,

title = "Structural characterization of potato protease inhibitor I (Cv. Bintje) after expression in Pichia pastoris",

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 -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",

keywords = "circular-dichroism, stability, proteins, chymotrypsin, tubers, solubility, elkana",

author = "{van den Broek}, L.A.M. and L.A.M. Pouvreau and G. Lommerse and B. Schipper and {van Koningsveld}, G.A. and H. Gruppen",

year = "2004",

doi = "10.1021/jf049832x",

language = "English",

volume = "52",

pages = "4928--4934",

journal = "Journal of Agricultural and Food Chemistry",

issn = "0021-8561",

publisher = "American Chemical Society",

}

TY - JOUR

T1 - Structural characterization of potato protease inhibitor I (Cv. Bintje) after expression in Pichia pastoris

AU - van den Broek, L.A.M.

AU - Pouvreau, L.A.M.

AU - Lommerse, G.

AU - Schipper, B.

AU - van Koningsveld, G.A.

AU - Gruppen, H.

PY - 2004

Y1 - 2004

N2 - 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 -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

AB - 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 -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

KW - circular-dichroism

KW - stability

KW - proteins

KW - chymotrypsin

KW - tubers

KW - solubility

KW - elkana

U2 - 10.1021/jf049832x

DO - 10.1021/jf049832x

M3 - Article

SN - 0021-8561

VL - 52

SP - 4928

EP - 4934

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

ER -

Structural characterization of potato protease inhibitor I (Cv. Bintje) after expression in Pichia pastoris

Abstract

Keywords

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