Molecular cloning and expression of subunit 12: a non-MCP and non-ATPase subunit of the 26 S protease.
A cDNA encoding subunit 12 (S12) of human erythrocyte 26 S protease has been isolated, sequenced and expressed. The cDNA contains an open reading frame that encodes a 36.6 kDA protein 96% identical to mouse Mov-34 and 67% identical to its Drosophila melanogaster homolog. Based on the high degree of sequence identity between human S12, mouse and Drosophila Mov-34 proteins, we conclude that the Mov-34 gene product is a component of the 26 S protease. Antibodies produced against two S12 fragments, Met1-Tyr95 (S12f95) and Met1-Leu205 (S12f205), react with S12 transferred to nitrocellulose from SDS-PAGE. In contrast, after transfer from native gels, the epitope(s) recognized by anti-S12f205 is exposed in the regulatory complex but appears to be masked when the regulatory complex associates with the multicatalytic protease.
Dubiel
W
W
Institut für Biochemie, Universitätsklinikum Charité, Humboldt-Universität, Berlin, Germany.
Ferrell
K
K
Dumdey
R
R
Standera
S
S
Prehn
S
S
Rechsteiner
M
M
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D004331:Drosophila melanogaster; D004912:Erythrocytes / Q000201:enzymology*; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010447:Peptide Hydrolases / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction; D046988:Proteasome Endopeptidase Complex; D021122:Protein Subunits / Q000737:chemistry; D011506:Proteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins / Q000737:chemistry; D012191:Retroviridae Proteins / Q000737:chemistry; D016415:Sequence Alignment; D017385:Sequence Homology
D018076:DNA, Complementary; D047908:Intracellular Signaling Peptides and Proteins; C488765:PSMD7 protein, human; D021122:Protein Subunits; D011506:Proteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D012191:Retroviridae Proteins; C072621:Psmd7 protein, mouse; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease