Expression of pace in host cells and methods of use thereof
AssigneesGenetics Institute, Inc., Chiron Corporation
InventorsPhilip J. Barr, Anthony J. Brake, Randal J. Kaufman, Patricia Tekamp-Olson, Louise Wasley, Polly A. Wong
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Abstract
Compositions and methods are provided for endopeptidase production and for enhanced efficiencies of processing heterologous precursor polypeptides to mature polypeptides, including proteins requiring gamma-carboxylation for biological activity. These compositions and methods utilize recombinant PACE, a mammalian endopeptidase that is specific for dibasic amino acid sites.
Claims14
A soluble endopeptidase protein precursor processing enzyme comprising the polypeptide sequence of SEQ ID NO:
- 2 or SEQ ID, NO:
- 4 which is truncated at between about amino acid #716 and amino acid #738 thereof, said protein having the biological activity of cleaving, at basic amino acid pairs, precursor polypeptides that require gamma-carboxylation for biological activity.
A soluble endopeptidase protein precursor processing enzyme comprising a truncated polypeptide sequence of SEQ ID NO:
- 2 or SEQ ID NO:
- 4, said truncation occurring at between about amino acid #716 and amino acid #738 thereof, said enzyme having the biological activity of cleaving, at basic amino acid pairs, precursor polypeptides that require gamma-carboxylation for biological activity said enzyme encoded by a nucleotide sequence of SEQ ID NO:
- 1 or SEQ ID NO:
- 3 modified by 1 to 4 codon changes.
A soluble endopeptidase protein precursor processing enzyme encoded by a nucleotide sequence of SEQ ID NO:
- 1 or SEQ ID NO:
- 3 truncated between about nucleotide #2553 and about nucleotide #2621, which sequence is modified by 1 to 4 codon changes which permit retention of the native enzyme activity of said enzyme capable of cleaving at basic amino acid pairs precursor polypeptides that require gamma-carboxylation for biological activity.
Dependencies are inferred from the claim text.