Isolation and cDNA Cloning of a Novel Galanin-like Peptide (GALP) from Porcine Hypothalamus

Abstract Galanin is a widely distributed neuropeptide with a variety of physiological functions. Three galanin receptor subtypes, GALR1, GALR2, and GALR3, have been reported. We isolated a novel galanin-like peptide (GALP) from porcine hypothalamus by observing its activity for increasing [35S]GTPγS binding to a membrane preparation of GALR2-transfected cells. The peptide had 60 amino acid residues and a non-amidated C terminus. The amino acid sequence of GALP-(9–21) was completely identical to that of galanin-(1–13). A cloned porcine GALP cDNA indicated that GALP was processed from a 120-amino acid GALP precursor protein. The structures of rat and human GALP-(1–60) were deduced from cloned cDNA, which indicated that the amino acid sequences 1–24 and 41–53 were highly conserved between humans, rats, and pigs. Receptor binding studies revealed that porcine GALP-(1–60) had a high affinity for the GALR2 receptor (IC50 = 0.24 nm) and a lower affinity for the GALR1 receptor (IC50 = 4.3 nm). In contrast, galanin showed high affinity for the GALR1 (IC50= 0.097 nm) and GALR2 receptors (IC50 = 0.48 nm). GALP is therefore an endogenous ligand that preferentially binds the GALR2 receptor, whereas galanin is relatively non-selective.