Frequency-domain measurements of the rotational dynamics of the tyrosine groups of calmodulin

Ignacy Gryczynski, J. R. Lakowicz, R. F. Steiner

Research output: Contribution to journalArticle

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Abstract

We used frequency-domain fluorometry to determine the intensity and anisotropy decay kinetics of tyrosine residues in calmodulin and its fragments. Excitation was provided by a continuous ultraviolet laser source, a frequency-doubled rhodamine 6G ring dye laser, whose output was externally modulated to 200 MHz. Both the intensity and anisotropy decays were found to be multiexponential and dependent upon temperature and solution conditions. By examination of calmodulin fragments we determined that energy transfer between the two tyrosine residues reduces the steady-state anisotropy values by about 20%. Additionally, the frequency-domain anisotropy decays indicate local torsional motions of the tyrosine residues, as well as significant individual motions of the two domains of calmodulin.

Original languageEnglish
Pages (from-to)49-59
Number of pages11
JournalBiophysical Chemistry
Volume30
Issue number1
DOIs
Publication statusPublished - 1 Jan 1988

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Keywords

  • Calmodulin
  • Fluorescence anisotropy decay
  • Fluorescence intensity decay
  • Frequency-domain fluorometry
  • Tyrosine

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