title: The generation of valosin-like peptides from a precursor protein in vitro as an extraction artifact.
abstract: Valosin is a 25 amino acid peptide recently isolated from the porcine gastrointestinal tract. The molecular forms of valosin-like immunoreactivity (VLIR) were examined following different tissue extraction procedures. Fractionation of tissue extracted with cold 0.1 M sodium hydroxide by Sephadex G50 gel permeation chromatography revealed a large form of VLIR (Kav = 0). Smaller forms of VLIR, Kav = 0.36 and 0.57 were obtained in tissue extracted by boiling in 0.5 M acetic acid. Acidification and boiling of the 0.1 M sodium hydroxide tissue extracts also generated smaller forms of VLIR of Kav = 0.36 and 0.57. Partially purified preparations of the large forms of VLIR extracted with sodium hydroxide could be disrupted into a smaller form of Kav = 0.57 by acidification and boiling. This smaller molecular form co-eluted with the synthetic 25 amino acid valosin standard. We conclude that valosin does not occur naturally but is an artifact generated by cleavage of a larger protein precursor upon acid extraction of tissues. Workers should be aware of the need to verify their extraction procedures when characterising novel peptides to avoid potential pitfalls such as acid/thermal cleavage of proteins.
authors.affiliation: Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London.
authors.forename: J S
authors.initials: JS
authors.lastname: Gill
authors.forename: M A
authors.initials: MA
authors.lastname: Ghatei
authors.forename: J
authors.initials: J
authors.lastname: Domin
authors.forename: S R
authors.initials: SR
authors.lastname: Bloom
mesh_terms: D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006358:Hot Temperature; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides / Q000302:isolation & purification*; D011498:Protein Precursors / Q000378:metabolism*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D011498:Protein Precursors; C046951:valosin