Yeast KEX2 endopeptidase correctly cleaves a neuroendocrine prohormone in mammalian cells |
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Authors: | G Thomas B A Thorne L Thomas R G Allen D E Hruby R Fuller J Thorner |
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Institution: | Vollum Institute for Advanced Biomedical Research, Oregon Health Sciences University, Portland 97201. |
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Abstract: | Mammalian cell lines (BSC-40, NG108-15, and GH4C1) that cannot process the murine neuroendocrine peptide precursor prepro-opiomelanocortin (mPOMC) when its synthesis is directed by a vaccinia virus vector were coinfected with a second recombinant vaccinia virus carrying the yeast KEX2 gene, which encodes an endopeptidase that cleaves at pairs of basic amino acid residues. mPOMC was cleaved intracellularly to a set of product peptides normally found in vivo, including mature gamma-lipotropin and beta-endorphin1-31. In GH4C1 cells (a rat pituitary line), product peptides were incorporated into stored secretory granules. These results suggest that the inability of any particular cell line to process a prohormone precursor is due to the absence of a suitable endogenous processing enzyme. |
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