Molecular cloning and expression of subunit 9 of the 26S proteasome

FEBS Lett. 1997 Mar 10;404(2-3):179-84. doi: 10.1016/s0014-5793(97)00126-9.

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

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • Caenorhabditis elegans / enzymology
  • Cloning, Molecular
  • Escherichia coli
  • Humans
  • Macromolecular Substances
  • Molecular Sequence Data
  • Open Reading Frames
  • Peptide Hydrolases / biosynthesis*
  • Peptide Hydrolases / chemistry*
  • Peptide Hydrolases / genetics
  • Proteasome Endopeptidase Complex*
  • Rabbits
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Saccharomyces cerevisiae / enzymology
  • Sequence Homology, Amino Acid
  • Sequence Tagged Sites

Substances

  • Macromolecular Substances
  • Recombinant Proteins
  • Peptide Hydrolases
  • Proteasome Endopeptidase Complex
  • ATP dependent 26S protease

Associated data

  • GENBANK/AF001212
  • GENBANK/X95644