Thermodynamics and kinetics of the pressure unfolding of phosphoglycerate kinase

Szabolcs Osváth, Manh Quynh Luu, L. Smeller

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Due to the relationship between compressibility and volume fluctuations, high-pressure studies provide vital insight into protein dynamics and function. Most high-pressure experiments were performed on small and fast folding proteins or model peptides. Here we show that a detailed kinetic study is necessary to extract reliable information from the high-pressure-induced structural conversion of large, slowly folding proteins. The pressure-jump unfolding kinetics of yeast phosphoglycerate kinase was recorded at pressures between 50 and 150 MPa. The time dependence of the conformational state of the protein was followed by tryptophan fluorescence measurements from 30 s to 2 h. The observed changes were described by a three-state model, and the volume change and the activation volume as well as the midpoint pressure of the transitions between the folded, intermediate, and unfolded states were determined. An interesting feature of the pressure unfolding of phosphoglycerate kinase was that the unfolding process speeds up with increasing pressure, which is the consequence of negative activation volumes for the folded → intermediate, intermediate → unfolded, and unfolded → intermediate transitions.

Original languageEnglish
Pages (from-to)10146-10150
Number of pages5
JournalBiochemistry
Volume48
Issue number42
DOIs
Publication statusPublished - Oct 27 2009

Fingerprint

Phosphoglycerate Kinase
Thermodynamics
Pressure
Kinetics
Protein folding
Protein Folding
Chemical activation
Compressibility
Tryptophan
Yeast
Proteins
Yeasts
Fluorescence
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Thermodynamics and kinetics of the pressure unfolding of phosphoglycerate kinase. / Osváth, Szabolcs; Luu, Manh Quynh; Smeller, L.

In: Biochemistry, Vol. 48, No. 42, 27.10.2009, p. 10146-10150.

Research output: Contribution to journalArticle

Osváth, Szabolcs ; Luu, Manh Quynh ; Smeller, L. / Thermodynamics and kinetics of the pressure unfolding of phosphoglycerate kinase. In: Biochemistry. 2009 ; Vol. 48, No. 42. pp. 10146-10150.
@article{9c6ce8b7c8284bcfb05b713086566b87,
title = "Thermodynamics and kinetics of the pressure unfolding of phosphoglycerate kinase",
abstract = "Due to the relationship between compressibility and volume fluctuations, high-pressure studies provide vital insight into protein dynamics and function. Most high-pressure experiments were performed on small and fast folding proteins or model peptides. Here we show that a detailed kinetic study is necessary to extract reliable information from the high-pressure-induced structural conversion of large, slowly folding proteins. The pressure-jump unfolding kinetics of yeast phosphoglycerate kinase was recorded at pressures between 50 and 150 MPa. The time dependence of the conformational state of the protein was followed by tryptophan fluorescence measurements from 30 s to 2 h. The observed changes were described by a three-state model, and the volume change and the activation volume as well as the midpoint pressure of the transitions between the folded, intermediate, and unfolded states were determined. An interesting feature of the pressure unfolding of phosphoglycerate kinase was that the unfolding process speeds up with increasing pressure, which is the consequence of negative activation volumes for the folded → intermediate, intermediate → unfolded, and unfolded → intermediate transitions.",
author = "Szabolcs Osv{\'a}th and Luu, {Manh Quynh} and L. Smeller",
year = "2009",
month = "10",
day = "27",
doi = "10.1021/bi900922f",
language = "English",
volume = "48",
pages = "10146--10150",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "42",

}

TY - JOUR

T1 - Thermodynamics and kinetics of the pressure unfolding of phosphoglycerate kinase

AU - Osváth, Szabolcs

AU - Luu, Manh Quynh

AU - Smeller, L.

PY - 2009/10/27

Y1 - 2009/10/27

N2 - Due to the relationship between compressibility and volume fluctuations, high-pressure studies provide vital insight into protein dynamics and function. Most high-pressure experiments were performed on small and fast folding proteins or model peptides. Here we show that a detailed kinetic study is necessary to extract reliable information from the high-pressure-induced structural conversion of large, slowly folding proteins. The pressure-jump unfolding kinetics of yeast phosphoglycerate kinase was recorded at pressures between 50 and 150 MPa. The time dependence of the conformational state of the protein was followed by tryptophan fluorescence measurements from 30 s to 2 h. The observed changes were described by a three-state model, and the volume change and the activation volume as well as the midpoint pressure of the transitions between the folded, intermediate, and unfolded states were determined. An interesting feature of the pressure unfolding of phosphoglycerate kinase was that the unfolding process speeds up with increasing pressure, which is the consequence of negative activation volumes for the folded → intermediate, intermediate → unfolded, and unfolded → intermediate transitions.

AB - Due to the relationship between compressibility and volume fluctuations, high-pressure studies provide vital insight into protein dynamics and function. Most high-pressure experiments were performed on small and fast folding proteins or model peptides. Here we show that a detailed kinetic study is necessary to extract reliable information from the high-pressure-induced structural conversion of large, slowly folding proteins. The pressure-jump unfolding kinetics of yeast phosphoglycerate kinase was recorded at pressures between 50 and 150 MPa. The time dependence of the conformational state of the protein was followed by tryptophan fluorescence measurements from 30 s to 2 h. The observed changes were described by a three-state model, and the volume change and the activation volume as well as the midpoint pressure of the transitions between the folded, intermediate, and unfolded states were determined. An interesting feature of the pressure unfolding of phosphoglycerate kinase was that the unfolding process speeds up with increasing pressure, which is the consequence of negative activation volumes for the folded → intermediate, intermediate → unfolded, and unfolded → intermediate transitions.

UR - http://www.scopus.com/inward/record.url?scp=70350236636&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70350236636&partnerID=8YFLogxK

U2 - 10.1021/bi900922f

DO - 10.1021/bi900922f

M3 - Article

C2 - 19775155

AN - SCOPUS:70350236636

VL - 48

SP - 10146

EP - 10150

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 42

ER -