Structure and stability of the potato cysteine protease inhibitor group (Cv. Elkana)

L.A.M. Pouvreau, C.H.M. Kroef, H. Gruppen, G.A. van Koningsveld, L.A.M. van den Broek, A.G.J. Voragen

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Scopus)

Abstract

The conformational stability of potato cysteine protease inhibitor (PCPI), the second most abundant protease inhibitor group in potato tuber, was investigated at ambient temperature and upon heating using far- and near-UV circular dichroism spectroscopy, fluorescence spectroscopy, and differential scanning calorimetry (DSC). The PCPI isoforms investigated have a highly similar structure at both the secondary and the tertiary level. PCPI isoforms show structural properties similar to those of the potato serine protease inhibitor group and the Kunitz type soybean trypsin inhibitor, a known -II protein. Therefore, PCPI isoforms are also classified as members of the -II protein subclass. Results show that the thermal unfolding of PCPI isoforms does not follow a two-state mechanism and that at least one intermediate is present. The occurrence of this intermediate is most apparent in the thermal unfolding of PCPI 8.3 as indicated by the presence of two peaks in the DSC thermogram. Additionally, the formation of aggregates (>100 kDa), especially at low scan rates, increases the apparent cooperativity of the unfolding
Original languageEnglish
Pages (from-to)5739-5746
JournalJournal of Agricultural and Food Chemistry
Volume53
Issue number14
DOIs
Publication statusPublished - 2005

Fingerprint

Cysteine Proteinase Inhibitors
cysteine proteinase inhibitors
Solanum tuberosum
potatoes
Protein Isoforms
Differential scanning calorimetry
Kunitz Soybean Trypsin Inhibitor
Differential Scanning Calorimetry
proteinase inhibitors
differential scanning calorimetry
heat
Circular dichroism spectroscopy
fluorescence emission spectroscopy
Serine Proteinase Inhibitors
Hot Temperature
Fluorescence spectroscopy
circular dichroism spectroscopy
Protease Inhibitors
Ultraviolet spectroscopy
Structural properties

Keywords

  • circular-dichroism spectra
  • molten-globule state
  • plasminogen-activator
  • proteinase-inhibitor
  • conformation
  • denaturation
  • fluorescence
  • spectroscopy
  • binding
  • family

Cite this

@article{73511fe59bd74af19f153342e9d7b83e,
title = "Structure and stability of the potato cysteine protease inhibitor group (Cv. Elkana)",
abstract = "The conformational stability of potato cysteine protease inhibitor (PCPI), the second most abundant protease inhibitor group in potato tuber, was investigated at ambient temperature and upon heating using far- and near-UV circular dichroism spectroscopy, fluorescence spectroscopy, and differential scanning calorimetry (DSC). The PCPI isoforms investigated have a highly similar structure at both the secondary and the tertiary level. PCPI isoforms show structural properties similar to those of the potato serine protease inhibitor group and the Kunitz type soybean trypsin inhibitor, a known -II protein. Therefore, PCPI isoforms are also classified as members of the -II protein subclass. Results show that the thermal unfolding of PCPI isoforms does not follow a two-state mechanism and that at least one intermediate is present. The occurrence of this intermediate is most apparent in the thermal unfolding of PCPI 8.3 as indicated by the presence of two peaks in the DSC thermogram. Additionally, the formation of aggregates (>100 kDa), especially at low scan rates, increases the apparent cooperativity of the unfolding",
keywords = "circular-dichroism spectra, molten-globule state, plasminogen-activator, proteinase-inhibitor, conformation, denaturation, fluorescence, spectroscopy, binding, family",
author = "L.A.M. Pouvreau and C.H.M. Kroef and H. Gruppen and {van Koningsveld}, G.A. and {van den Broek}, L.A.M. and A.G.J. Voragen",
year = "2005",
doi = "10.1021/jf050306v",
language = "English",
volume = "53",
pages = "5739--5746",
journal = "Journal of Agricultural and Food Chemistry",
issn = "0021-8561",
publisher = "American Chemical Society",
number = "14",

}

Structure and stability of the potato cysteine protease inhibitor group (Cv. Elkana). / Pouvreau, L.A.M.; Kroef, C.H.M.; Gruppen, H.; van Koningsveld, G.A.; van den Broek, L.A.M.; Voragen, A.G.J.

In: Journal of Agricultural and Food Chemistry, Vol. 53, No. 14, 2005, p. 5739-5746.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Structure and stability of the potato cysteine protease inhibitor group (Cv. Elkana)

AU - Pouvreau, L.A.M.

AU - Kroef, C.H.M.

AU - Gruppen, H.

AU - van Koningsveld, G.A.

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

AU - Voragen, A.G.J.

PY - 2005

Y1 - 2005

N2 - The conformational stability of potato cysteine protease inhibitor (PCPI), the second most abundant protease inhibitor group in potato tuber, was investigated at ambient temperature and upon heating using far- and near-UV circular dichroism spectroscopy, fluorescence spectroscopy, and differential scanning calorimetry (DSC). The PCPI isoforms investigated have a highly similar structure at both the secondary and the tertiary level. PCPI isoforms show structural properties similar to those of the potato serine protease inhibitor group and the Kunitz type soybean trypsin inhibitor, a known -II protein. Therefore, PCPI isoforms are also classified as members of the -II protein subclass. Results show that the thermal unfolding of PCPI isoforms does not follow a two-state mechanism and that at least one intermediate is present. The occurrence of this intermediate is most apparent in the thermal unfolding of PCPI 8.3 as indicated by the presence of two peaks in the DSC thermogram. Additionally, the formation of aggregates (>100 kDa), especially at low scan rates, increases the apparent cooperativity of the unfolding

AB - The conformational stability of potato cysteine protease inhibitor (PCPI), the second most abundant protease inhibitor group in potato tuber, was investigated at ambient temperature and upon heating using far- and near-UV circular dichroism spectroscopy, fluorescence spectroscopy, and differential scanning calorimetry (DSC). The PCPI isoforms investigated have a highly similar structure at both the secondary and the tertiary level. PCPI isoforms show structural properties similar to those of the potato serine protease inhibitor group and the Kunitz type soybean trypsin inhibitor, a known -II protein. Therefore, PCPI isoforms are also classified as members of the -II protein subclass. Results show that the thermal unfolding of PCPI isoforms does not follow a two-state mechanism and that at least one intermediate is present. The occurrence of this intermediate is most apparent in the thermal unfolding of PCPI 8.3 as indicated by the presence of two peaks in the DSC thermogram. Additionally, the formation of aggregates (>100 kDa), especially at low scan rates, increases the apparent cooperativity of the unfolding

KW - circular-dichroism spectra

KW - molten-globule state

KW - plasminogen-activator

KW - proteinase-inhibitor

KW - conformation

KW - denaturation

KW - fluorescence

KW - spectroscopy

KW - binding

KW - family

U2 - 10.1021/jf050306v

DO - 10.1021/jf050306v

M3 - Article

VL - 53

SP - 5739

EP - 5746

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

SN - 0021-8561

IS - 14

ER -