Peptide V: a VGF-derived neuropeptide purified from bovine posterior pituitary

• Published in: Endocrinology (Philadelphia) Vol. 135; no. 6; p. 2742
• Main Authors: Liu, J W, Andrews, P C, Mershon, J L, Yan, C, Allen, D L, Ben-Jonathan, N
• Format: Journal Article
• Language: English
• Published: United States 01.12.1994
• Subjects: Animals; Cattle; Chromatography, Gel; Chromatography, High Pressure Liquid; Mass Spectrometry; Neuropeptides - chemical synthesis; Neuropeptides - chemistry; Peptide Mapping; Pituitary Gland, Posterior - chemistry
• ISSN: 0013-7227
• DOI: 10.1210/endo.135.6.7988466

The objective of this study was to purify PRL-releasing factor (PRF) from the bovine posterior pituitary (PP) and determine its structure. Five hundred bovine PPs were acid extracted and fractionated using gel filtration chromatography followed by semipreparative and analytical HPLC. PRF activity was determined by an in vitro bioassay. After six chromatographic steps, a single peak with PRF activity was resolved. As determined by mass spectrometry and microsequencing, this peak contained a major peptide composed of 30 amino acids with a mol wt of 3708K. A synthetic peptide was then produced by solid-phase synthesis. When tested both in vivo and in vitro, the synthetic peptide lacked PRF activity. Further HPLC fractionation under different conditions resolved the synthetic peptide from a highly purified PRF activity. This indicated that the isolated peptide was coincidentally eluted with PRF during the purification. The major isolated peptide has 94% identity with a sequence at the C-terminus of a rat protein named VGF. VGF is a nerve growth factor-inducible protein that has been identified in PC12 cells and is localized in selected sites throughout the central nervous system. The isolated peptide has an Arg-Arg cleavage site at its junction within the VGF protein. Based on this information, we named this substance Peptide V (VGF-derived peptide). We postulate that Peptide V is: 1) a natural cleavage product of the VGF protein; 2) produced and processed either in the hypothalamus or within the pituitary proper, and 3) a releasable peptide that fulfills one or more endocrine functions.