Photoaffinity Labeling of RARs and Mapping of Labeled Sites by an Endoproteinase Combination Technique

  • Yuichi Hashimoto
  • Toru Sasaki
Part of the Methods in Molecular Biology book series (MIMB, volume 89)


Photoaffinity labeling and mapping of the labeled site(s) are a superior and convenient methodology for initial structural investigation of ligand-receptor complexes. The method we describe in this chapter consists of five steps.
  1. 1.

    Design and synthesis of a photoreactive probe for retinoic acid receptor (RAR)-labeling

  2. 2.

    Photoaffmlty labeling of RAR

  3. 3.

    Sequential digestion of the labeled RAR with endoproteinases

  4. 4

    High-performance liquid chromatography (HPLC) analysis of the digests to determine the amino-acid sequence context of the labeled sites (endoproteinase combination technique).

  5. 5.

    Mapping of the labeled site(s) by comparison of the determined context with the known amino-acid sequence of the RAR</lt>



Retinoic Acid Receptor Thionyl Chloride Fluorescent Peak Incubation Buffer Dialysis Buffer 
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Copyright information

© Humana Press Inc, Totowa, NJ 1998

Authors and Affiliations

  • Yuichi Hashimoto
    • 1
  • Toru Sasaki
    • 1
  1. 1.Institute of Molecular and Cellular BiosciencesUniversity of TokyoJapan

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