Microwave-assisted solid-phase peptide synthesis of neurosecretory protein GL composed of 80 amino acid residues

We recently identified a novel cDNA encoding a small secretory protein of 80 amino acid residues, termed neurosecretory protein GL (NPGL), from the chicken hypothalamus. Homologs of NPGL have been reported to be present in mammals, such as human and rat. NPGL is amidated at its C‐terminus, contains an intramolecular disulfide bond, and is hydrophobic in nature. In this study, we have optimized the synthesis of the entire 80‐amino acid peptide sequence of rat NPGL by microwave‐assisted solid‐phase peptide synthesis. NPGL was obtained with a 10% yield when the coupling reactions were performed using 1‐[Bis(dimethylamino)methylene]‐1H‐1,2,3‐triazolo[4,5‐b]pyridinium‐3‐oxid hexafluorophosphate (HATU) at 50 °C for 5 min, and Fmoc deprotections were performed using 40% piperidine containing 0.1 M HOBt. Furthermore, the disulfide bond of NPGL was formed with 20% yield with the use of glutathione‐containing redox buffer and 50% acetonitrile. Copyright © 2015 European Peptide Society and John Wiley & Sons, Ltd. Neurosecretory protein GL (NPGL) is a novel hydrophobic peptide containing 80‐amino acid residues, an intramolecular disulfide bond, and an amidated C‐terminus. In this study, we synthesized rat NPGL by microwave‐assisted solid‐phase peptide synthesis. Amino acid couplings and Fmoc deprotections were performed using HATU at 50 °C for 5 min and 40% piperidine/0.1 M HOBt at 50 °C for 3 min, respectively. Then, intramolecular disulfide bond was formed using glutathione/50% acetonitrile at room temperature for 48 h.