Photoaffinity labelling of peptide hormone receptors

Photoreactive peptide derivatives for the labelling of hormone receptors are usually prepared by inserting a chemically stable aryl azide or nitroaryl azide into a specific site of the molecule, such as an alpha or omega amino or carboxyl group, or into the side-chain of an Arg, Cys, His, Trp or Tyr. With p-azidophenylalanine (Pap), a more or less isosteric replacement of Tyr or Phe can be achieved when other alterations would impair the biological activity of the hormone. Reversible attachment of photoreactive groups via S-S linkage to, e.g., SH-Trp makes it possible to release the covalently bound hormone from the receptor. This is an advantage when photolabelling is used for the isolation of receptors. Photoaffinity labelling of intact cell systems may reveal insights into dynamic aspects, such as receptor inactivation and turnover or the temporal involvement of second messengers, and may be particularly useful for the study of peptide action on target cells occurring only in small numbers. The Xenopus MSH-melanophore system which is stimulated irreversibly upon UV-irradiation of photoreactive alpha-MSH represents such an example.