Structural changes associated with the acute thermal instability of Rubisco activase

Csengele Barta, Alison M. Dunkle, Rebekka Wachter, Michael E. Salvucci

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Inhibition of photosynthesis by heat has been linked to the instability of the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) chaperone, Rubisco activase. Examination of the recombinant enzyme showed that ADP and ATP protected against inactivation, whereas Mg2+ promoted inactivation. Heating caused aggregation of Rubisco activase characterized by disruption of secondary structure content and formation of insoluble protein. In contrast, incubation at room temperature without nucleotide caused the active ∼660kDa protein to form a soluble, but inactive aggregate of >2×106Da. Circular dichroism (CD) spectroscopy and fluorescence established that structural perturbations in the aggregate did not reduce alpha-helical content significantly. Differences in the thermal stability between wild type and mutant Rubisco activase were observed for the recombinant proteins and when the proteins were expressed in transgenic Arabidopsis. That the sensitivity of these plants to heat differs indicates that the thermal instability of Rubisco activase is a main determinant of the temperature-sensitivity of photosynthesis.

Original languageEnglish (US)
Pages (from-to)17-25
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume499
Issue number1-2
DOIs
StatePublished - Jul 2010

Fingerprint

Oxygenases
Tissue Plasminogen Activator
Hot Temperature
Photosynthesis
Circular dichroism spectroscopy
Proteins
Temperature
Fluorescence Spectrometry
Circular Dichroism
Recombinant Proteins
Arabidopsis
Heating
Adenosine Diphosphate
Thermodynamic stability
Agglomeration
Nucleotides
Adenosine Triphosphate
Fluorescence
ribulose-1,5 diphosphate
Enzymes

Keywords

  • Adenine nucleotides
  • Calvin cycle
  • Heat stress
  • Magnesium ions
  • Photosynthetic CO fixation
  • Thermal stability

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Structural changes associated with the acute thermal instability of Rubisco activase. / Barta, Csengele; Dunkle, Alison M.; Wachter, Rebekka; Salvucci, Michael E.

In: Archives of Biochemistry and Biophysics, Vol. 499, No. 1-2, 07.2010, p. 17-25.

Research output: Contribution to journalArticle

Barta, Csengele ; Dunkle, Alison M. ; Wachter, Rebekka ; Salvucci, Michael E. / Structural changes associated with the acute thermal instability of Rubisco activase. In: Archives of Biochemistry and Biophysics. 2010 ; Vol. 499, No. 1-2. pp. 17-25.
@article{57049dd16ad6474aaf5eb3b890eedce7,
title = "Structural changes associated with the acute thermal instability of Rubisco activase",
abstract = "Inhibition of photosynthesis by heat has been linked to the instability of the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) chaperone, Rubisco activase. Examination of the recombinant enzyme showed that ADP and ATP protected against inactivation, whereas Mg2+ promoted inactivation. Heating caused aggregation of Rubisco activase characterized by disruption of secondary structure content and formation of insoluble protein. In contrast, incubation at room temperature without nucleotide caused the active ∼660kDa protein to form a soluble, but inactive aggregate of >2×106Da. Circular dichroism (CD) spectroscopy and fluorescence established that structural perturbations in the aggregate did not reduce alpha-helical content significantly. Differences in the thermal stability between wild type and mutant Rubisco activase were observed for the recombinant proteins and when the proteins were expressed in transgenic Arabidopsis. That the sensitivity of these plants to heat differs indicates that the thermal instability of Rubisco activase is a main determinant of the temperature-sensitivity of photosynthesis.",
keywords = "Adenine nucleotides, Calvin cycle, Heat stress, Magnesium ions, Photosynthetic CO fixation, Thermal stability",
author = "Csengele Barta and Dunkle, {Alison M.} and Rebekka Wachter and Salvucci, {Michael E.}",
year = "2010",
month = "7",
doi = "10.1016/j.abb.2010.04.022",
language = "English (US)",
volume = "499",
pages = "17--25",
journal = "Archives of Biochemistry and Biophysics",
issn = "0003-9861",
publisher = "Academic Press Inc.",
number = "1-2",

}

TY - JOUR

T1 - Structural changes associated with the acute thermal instability of Rubisco activase

AU - Barta, Csengele

AU - Dunkle, Alison M.

AU - Wachter, Rebekka

AU - Salvucci, Michael E.

PY - 2010/7

Y1 - 2010/7

N2 - Inhibition of photosynthesis by heat has been linked to the instability of the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) chaperone, Rubisco activase. Examination of the recombinant enzyme showed that ADP and ATP protected against inactivation, whereas Mg2+ promoted inactivation. Heating caused aggregation of Rubisco activase characterized by disruption of secondary structure content and formation of insoluble protein. In contrast, incubation at room temperature without nucleotide caused the active ∼660kDa protein to form a soluble, but inactive aggregate of >2×106Da. Circular dichroism (CD) spectroscopy and fluorescence established that structural perturbations in the aggregate did not reduce alpha-helical content significantly. Differences in the thermal stability between wild type and mutant Rubisco activase were observed for the recombinant proteins and when the proteins were expressed in transgenic Arabidopsis. That the sensitivity of these plants to heat differs indicates that the thermal instability of Rubisco activase is a main determinant of the temperature-sensitivity of photosynthesis.

AB - Inhibition of photosynthesis by heat has been linked to the instability of the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) chaperone, Rubisco activase. Examination of the recombinant enzyme showed that ADP and ATP protected against inactivation, whereas Mg2+ promoted inactivation. Heating caused aggregation of Rubisco activase characterized by disruption of secondary structure content and formation of insoluble protein. In contrast, incubation at room temperature without nucleotide caused the active ∼660kDa protein to form a soluble, but inactive aggregate of >2×106Da. Circular dichroism (CD) spectroscopy and fluorescence established that structural perturbations in the aggregate did not reduce alpha-helical content significantly. Differences in the thermal stability between wild type and mutant Rubisco activase were observed for the recombinant proteins and when the proteins were expressed in transgenic Arabidopsis. That the sensitivity of these plants to heat differs indicates that the thermal instability of Rubisco activase is a main determinant of the temperature-sensitivity of photosynthesis.

KW - Adenine nucleotides

KW - Calvin cycle

KW - Heat stress

KW - Magnesium ions

KW - Photosynthetic CO fixation

KW - Thermal stability

UR - http://www.scopus.com/inward/record.url?scp=77953722515&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77953722515&partnerID=8YFLogxK

U2 - 10.1016/j.abb.2010.04.022

DO - 10.1016/j.abb.2010.04.022

M3 - Article

C2 - 20450882

AN - SCOPUS:77953722515

VL - 499

SP - 17

EP - 25

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

IS - 1-2

ER -