Transformation of an irreversible MSH antagonist into an irreversible MSH agonist by differential receptor crosslinking using the photo-affinity technique.

Photocrosslinking of receptors for alpha-melanocyte-stimulating hormone (alpha-MSH) on melanophores of frogs and lizards has been shown to induce long-lasting receptor stimulation whereby the photoreactive alpha-MSH may contain one or two photolabels in positions 1, 7, 9, or 13. The chemical synthesis and biological testing of an alpha-MSH analogue is now described which contains three photoreactive groups in positions 1, 9 and 13, one of which with a cleavable S-S disulphide bridge: [ApSSpr-Ser1, Trp(Naps)9 Pap13]-alpha-MSH. Photocrosslinking of MSH receptors on melanophores of Anolis carolinensis with this analogue led to almost complete receptor blockade which could be transformed into long-lasting receptor stimulation by exposure to a thiol reagent. By contrast, the analogue containing only two photoreactive groups in positions 9 and 13, [Trp(Naps)9, Pap13]-alpha-MSH, produced long-lasting receptor stimulation which was not altered by the thiol reagent. These results demonstrate that one and the same peptide ligand may contain structural information for both receptor activation and inhibition and that the receptor may become arrested in an activated or inhibited state by multiple photocrosslinking, depending on the relative positions of these crosslinks.