Purification and Characterization of the Danaus Plexippus Cryptochromes

Abstract

Cryptochromes are flavoproteins that function as blue-light photoreceptors to regulate growth in plants and to entrain the circadian clock in animals. The photochemical and signaling mechanisms by which cryptochromes function are still undefined. Two cryptochromes identified in Danaus plexippus have shown distinct in vivo activities where DpCryl is similar to Drosophila melanogaster cryptochrome and DpCry2 is similar to mammalian cryptochromes. I purified and biochemically characterized both D. plexippus cryptochromes. DpCryl is the first animal cryptochrome to contain near-stoichiometric flavin. Photoreduction of FADox to FAD and fluorescence lifetime measurements showing an extremely short lifetime of flavin in DpCryl both suggest a novel photochemical mechanism for DpCryl. DpCry2 contained only trace amounts of flavin and both proteins contained sub-stoichiometric MTHF. Purification of DpCryl as the first animal cryptochrome with near-stoichiometric flavin will enable crucial in vitro biochemical and biophysical experiments that have never been conducted on an animal cryptochrome.

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Document Details

Document Type
Technical Report
Publication Date
Jan 01, 2006
Accession Number
ADA462912

Entities

People

  • Tracy R. Denaro

Organizations

  • University of North Carolina at Chapel Hill

Tags

Communities of Interest

  • Biomedical

DTIC Thesaurus Topics

  • Absorption Spectra
  • Air Force
  • Amino Acids
  • Animals
  • Cells
  • Chemical Reactions
  • Chemistry
  • Crystal Structure
  • Electron Transfer
  • Energy Transfer
  • Lasers
  • Lepidoptera
  • Photochemical Reactions
  • Proteins
  • Recombinant Proteins
  • Spectroscopy
  • Transcription Factors

Fields of Study

  • Biology
  • Computer science

Readers

  • Marine Propulsion Engineering and Naval Architecture
  • Molecular and Cellular Biochemistry
  • Munitions and Ordnance Engineering

Technology Areas

  • Biotechnology