Identification and characterization of desacyl-ghrelin

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Ghrelin is a peptide hormone to regulate appetite and is known to have two forms (acyl ghrelin and des-acyl ghrelin). Ghrelin receptor (GHSR), which recognizes the post-translational acyl-modification, expressed in various tissues. However, ghrelin without lipid-modification does not bind to GHSR and functions antagonistic to acyl ghrelin. These findings suggested the presenceof unknown receptor specific for des-acyl ghrelin. We performed affinity-purification of des-acyl ghrelin-binding protein using pig brain extract. We tried to elute des-acyl ghrelin-associated protein by high salt condition (1M NaCl) but detected no significant condensation of candidate molecules. Next, we performed conventional receptor panning assay using COS7 cells transfected with sub-pooled mouse brain cDNA libraries. We detected several pools as “positive pools” and further selection of procedures for cloning is in process.