Abstract
Processing of rat prodynorphin (proDyn) by the mouse prohormone convertase PC1 was investigated. Recombinant vaccinia virus vectors were used to coexpress proDyn and PC1 in rat PC12 pheochromocytoma and mouse AtT-20 corticotroph cells. In vitro experiments were also conducted by co-incubating purified proDyn and PC1. The results demonstrate that PC1 cleaves proDyn at pairs of basic residues to yield 10 and 16 kDa high molecular weight (HMW) intermediates. Additionally, PC1 cleaves proDyn at a single arginine residue to yield an 8 kDa product and the C-peptide. This demonstrates that PC1 cleaves proDyn at single and pairs of basic residues.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Arginine*
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Aspartic Acid Endopeptidases / chemistry
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Aspartic Acid Endopeptidases / genetics
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Aspartic Acid Endopeptidases / metabolism*
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Binding Sites
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C-Peptide / analysis
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C-Peptide / metabolism
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CHO Cells
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Cell Line
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Cricetinae
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Enkephalins / chemistry
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Enkephalins / genetics
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Enkephalins / metabolism*
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Genetic Vectors
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Molecular Sequence Data
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Molecular Weight
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PC12 Cells
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Proprotein Convertase 1*
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Proprotein Convertases
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Protein Precursors / chemistry
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Protein Precursors / genetics
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Protein Precursors / metabolism*
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Rats
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Transfection
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Vaccinia virus / genetics
Substances
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C-Peptide
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Enkephalins
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Protein Precursors
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Recombinant Proteins
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preproenkephalin
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Arginine
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Proprotein Convertases
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Pcsk1 protein, mouse
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Proprotein Convertase 1
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Aspartic Acid Endopeptidases