title: Expression of recombinant pro-neuropeptide Y, proopiomelanocortin, and proenkephalin: relative processing by 'prohormone thiol protease' (PTP).
abstract: The preference of the 'prohormone thiol protease' (PTP), a candidate prohormone processing enzyme, for different peptide precursors was assessed in vitro with recombinant prohormones near estimated in vivo levels. Pro-neuropeptide Y (pro-NPY), proopiomelanocortin (POMC), and proenkephalin (PE) were expressed at high levels in E. coli. Purification of prohormones utilized a combination of DEAE-Sepharose, Mono Q, and preparative electrophoresis. PTP cleaved PE most readily, and also cleaved pro-NPY. The processing of POMC by PTP was minimal. These results demonstrate PTP's preference for certain prohormone substrates.
authors.affiliation: Department of Biochemistry, Uniformed Services University, Bethesda, MD, USA.
authors.forename: M R
authors.initials: MR
authors.lastname: Schiller
authors.forename: A B
authors.initials: AB
authors.lastname: Kohn
authors.forename: L M
authors.initials: LM
authors.lastname: Mende-Muelller
authors.forename: K
authors.initials: K
authors.lastname: Miller
authors.forename: V Y
authors.initials: VY
authors.lastname: Hook
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004745:Enkephalins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009478:Neuropeptide Y / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011333:Pro-Opiomelanocortin / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013552:Swine
chemical_list: D004745:Enkephalins; D009478:Neuropeptide Y; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C029992:proenkephalin; C081158:proneuropeptide Y; D011333:Pro-Opiomelanocortin; D003546:Cysteine Endopeptidases; C083275:prohormone thiol protease