Expression of recombinant pro-neuropeptide Y, proopiomelanocortin, and proenkephalin: relative processing by 'prohormone thiol protease' (PTP).
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.
Amino acid sequence; Animals; Base Sequence; Biosynthesis; Cysteine Endopeptidases/metabolism; Endopeptidases; Enkephalins; Enkephalins/biosynthesis; Enkephalins/genetics; Enkephalins/isolation & purification; Enkephalins/metabolism; Escherichia coli; Escherichia coli/genetics; Genetics; Molecular Sequence Data; Nucleotide sequence; Neuropeptide Y; Neuropeptide Y/biosynthesis; Neuropeptide Y/genetics; Neuropeptide Y/isolation & purification; Neuropeptide Y/metabolism; Proopiomelanocortin; Pro-Opiomelanocortin/biosynthesis; Pro-Opiomelanocortin/genetics; Pro-Opiomelanocortin/isolation & purification; Pro-Opiomelanocortin/metabolism; Proprotein convertases; Protein precursors; Protein Precursors/biosynthesis; Protein Precursors/genetics; Protein Precursors/isolation & purification; Protein Precursors/metabolism; Protein Processing; Post-Translational; Proteins—Synthesis; Rats; Recombinant Fusion Proteins/biosynthesis; Recombinant Fusion Proteins/isolation & purification; Recombinant Fusion Proteins/metabolism; Substrate Specificity; Swine
Biochemistry | Life Sciences | Medicine and Health Sciences | Molecular Biology
Schiller, M. R.,
Kohn, A. B.,
Mende-Mueller, L. M.,
Hook, V. Y.
Expression of recombinant pro-neuropeptide Y, proopiomelanocortin, and proenkephalin: relative processing by 'prohormone thiol protease' (PTP)..
FEBS Letters, 382(1-2),