Molecular cloning and expression of subunit 9 of the 26S proteasome.
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.
Hoffman
L
L
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City 84132, USA. lhoffman@msscc.med.utah.edu
Rechsteiner
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D017173:Caenorhabditis elegans / Q000201:enzymology; D003001:Cloning, Molecular; D004926:Escherichia coli; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010447:Peptide Hydrolases / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D046988:Proteasome Endopeptidase Complex; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D017386:Sequence Homology, Amino Acid; D016324:Sequence Tagged Sites
D046911:Macromolecular Substances; D011994:Recombinant Proteins; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease