Target-peptide-induced changes in the structure and dynamics of calmodulin as probed by frequency domain fluorimetry of bound Tb(III)

Patricia B. O'Hara, Mohammad A. Rahman, Anastasia Rowland, A. John Turjoman

Research output: Contribution to journalArticle

Abstract

Tb(III) luminescence is used to probe the conformational change induced in the calcium regulatory protein calmodulin upon binding to a target peptide. The luminescence lifetime for Tb(III) measured by frequency domain fluorimetry increases from 1278 μs for the calmodulin complex to 1496 μs for the complex of calmodulin and M13, a peptide derived from the calmodulin target protein myosin light chain kinase. The intensity of the Tb(III) emission increases over the solution value by a factor of 726 and 891 when bound to calmodulin and to the complex of calmodulin and M13 respectively. The sensitivity of the Tb(III) decay rate to deuterated solvent was also measured and is consistent with a single water molecule ound to the metal in both the calmodulin and calmodulin-M13 complex. The most dramatic change induced by M13 is the threefold reduction in the width of the Tb(III) lifetime distribution, which is interpreted to be a target-peptide-induced annealing of the previously flexible metal-binding site.

Original languageEnglish (US)
Pages (from-to)15-21
Number of pages7
JournalJournal of Photochemistry and Photobiology B: Biology
Volume30
Issue number1
DOIs
StatePublished - 1995
Externally publishedYes

Fingerprint

Calmodulin
calmodulin
Fluorometry
fluorometry
Peptides
peptides
Luminescence
luminescence
Metals
Calmodulin-Binding Proteins
Myosin-Light-Chain Kinase
myosin light chain kinase
metals
proteins
Proteins
myosins
life (durability)
annealing
regulatory proteins
Protein Kinases

Keywords

  • Conformational change
  • Luminescence
  • M13
  • Rare earth ion

ASJC Scopus subject areas

  • Biophysics
  • Radiology Nuclear Medicine and imaging
  • Radiation
  • Radiological and Ultrasound Technology
  • Plant Science
  • Bioengineering
  • Physical and Theoretical Chemistry

Cite this

Target-peptide-induced changes in the structure and dynamics of calmodulin as probed by frequency domain fluorimetry of bound Tb(III). / O'Hara, Patricia B.; Rahman, Mohammad A.; Rowland, Anastasia; Turjoman, A. John.

In: Journal of Photochemistry and Photobiology B: Biology, Vol. 30, No. 1, 1995, p. 15-21.

Research output: Contribution to journalArticle

O'Hara, Patricia B. ; Rahman, Mohammad A. ; Rowland, Anastasia ; Turjoman, A. John. / Target-peptide-induced changes in the structure and dynamics of calmodulin as probed by frequency domain fluorimetry of bound Tb(III). In: Journal of Photochemistry and Photobiology B: Biology. 1995 ; Vol. 30, No. 1. pp. 15-21.
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AU - Rahman, Mohammad A.

AU - Rowland, Anastasia

AU - Turjoman, A. John

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N2 - Tb(III) luminescence is used to probe the conformational change induced in the calcium regulatory protein calmodulin upon binding to a target peptide. The luminescence lifetime for Tb(III) measured by frequency domain fluorimetry increases from 1278 μs for the calmodulin complex to 1496 μs for the complex of calmodulin and M13, a peptide derived from the calmodulin target protein myosin light chain kinase. The intensity of the Tb(III) emission increases over the solution value by a factor of 726 and 891 when bound to calmodulin and to the complex of calmodulin and M13 respectively. The sensitivity of the Tb(III) decay rate to deuterated solvent was also measured and is consistent with a single water molecule ound to the metal in both the calmodulin and calmodulin-M13 complex. The most dramatic change induced by M13 is the threefold reduction in the width of the Tb(III) lifetime distribution, which is interpreted to be a target-peptide-induced annealing of the previously flexible metal-binding site.

AB - Tb(III) luminescence is used to probe the conformational change induced in the calcium regulatory protein calmodulin upon binding to a target peptide. The luminescence lifetime for Tb(III) measured by frequency domain fluorimetry increases from 1278 μs for the calmodulin complex to 1496 μs for the complex of calmodulin and M13, a peptide derived from the calmodulin target protein myosin light chain kinase. The intensity of the Tb(III) emission increases over the solution value by a factor of 726 and 891 when bound to calmodulin and to the complex of calmodulin and M13 respectively. The sensitivity of the Tb(III) decay rate to deuterated solvent was also measured and is consistent with a single water molecule ound to the metal in both the calmodulin and calmodulin-M13 complex. The most dramatic change induced by M13 is the threefold reduction in the width of the Tb(III) lifetime distribution, which is interpreted to be a target-peptide-induced annealing of the previously flexible metal-binding site.

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