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Experientia

, Volume 39, Issue 10, pp 1063–1072 | Cite as

Reagents for photoaffinity labeling. I. Photobinding efficiency of aryl azido-, diazocyclopentadienyl-and ethyl diazomalonyl-derivatives of 9-aminoacridine

  • P. E. Nielsen
  • J. B. Hansen
  • T. Thomsen
  • O. Buchardt
Full Papers

Summary

A method for the comparison of photoaffinity labeling probes has been developed and tested with model reagents containing 5 different photoprobes attached to 9-acridinylamino groups through hexamethylenoxy or hexamethylenamino linkers. The fluorescence properties of the acridine part of the reagents were employed for detection of the labels. The merits and disadvantages of the different photoprobes are discussed. The photoreaction of the reagents with proteins (bovine serum albumin and histones), RNA (ribosomal), and DNA (calf thymus) were studied in order to compare the efficiency and suitability of aryl azido and diazo photoprobes. By using Pyrex-filtered light (λ>300 nm), it was observed that the reagents derived from 4-azidobenzoyl-(a), 4-azido-2-nitrophenyl- (b), or 2-diazopentadienylcarbonyl- (c) are the most efficient, labeling bovine serum albumin in yields of 22%, 9% and 9%, respectively, with relative rates of 0.25∶0.06∶1. The acridines containing photoprobesa, b andc were shown to function as photoaffinity labeling reagents of the histones in chromatin.

Keywords

Ethyl Albumin Bovine Serum Bovine Serum Albumin Serum Albumin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Birkhäuser Verlag 1983

Authors and Affiliations

  • P. E. Nielsen
    • 1
    • 2
  • J. B. Hansen
    • 1
    • 2
  • T. Thomsen
    • 1
    • 2
  • O. Buchardt
    • 1
    • 2
  1. 1.Department of Biochemistry B, The Panum InstituteUniversity of CopenhagenCopenhagen(Denmark)
  2. 2.Chemical Laboratory II, The H. C. Ørsted InstituteUniversity of CopenhagenCopenhagen(Denmark)

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