Abstract
Seven peptides from subunit 9 (S9) of the human 26S proteasome were sequenced and this information was used to clone a HeLa cDNA that encodes the 46 kDa subunit. Rabbit polyclonal antisera were made against a ubiquitin fusion protein containing 12 amino acids from S9 and against a full-length S9 expressed in E. coli. Western blot analysis showed that the S9-specific antibodies bound the 26S proteasome and its regulatory complex separated on non-denaturing gels. In SDS-PAGE samples of the two complexes, the S9-specific antibodies bound a single 46 kDa subunit. Thus, a cDNA encoding a novel 26S protease subunit has been isolated, sequenced, and expressed.
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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Base Sequence
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Blotting, Western
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Caenorhabditis elegans / enzymology
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Cloning, Molecular
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Escherichia coli
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Humans
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Macromolecular Substances
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Molecular Sequence Data
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Open Reading Frames
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Peptide Hydrolases / biosynthesis*
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Peptide Hydrolases / chemistry*
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Peptide Hydrolases / genetics
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Proteasome Endopeptidase Complex*
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Rabbits
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Saccharomyces cerevisiae / enzymology
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Sequence Homology, Amino Acid
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Sequence Tagged Sites
Substances
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Macromolecular Substances
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Recombinant Proteins
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Peptide Hydrolases
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Proteasome Endopeptidase Complex
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ATP dependent 26S protease
Associated data
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GENBANK/AF001212
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GENBANK/X95644