Ultrastructure of the approximately 26S complex containing the approximately 20S cylinder particle (multicatalytic proteinase/proteasome).
We have isolated a large protein complex of approximately 26S from Xenopus laevis oocytes and eggs which is composed of the approximately 20S cylinder particle (multicatalytic proteinase/proteasome) and additional proteinaceous components. In its polypeptide composition and sedimentation coefficient this approximately 26S complex closely resembles the 26S ubiquitin-dependent protease, a high molecular weight multienzyme complex recently described in the literature. Specific antibodies directed against a single subunit of the approximately 20S cylinder particle retain, on affinity columns, the large approximately 26S complex, and on sucrose gradients up to approximately 50% of the approximately 20S cylinder particles present in oocyte extracts sedimented with approximately 26S, suggesting that a large proportion of the approximately 20S particles exists in the cell as a component of the approximately 26S complex. Electron microscopy reveals the approximately 26S complex to be a symmetrical elongated macromolecular assembly of at least three protein particles. The central core of the complex is formed by the approximately 20S cylinder particle to which two other large components are attached at the ends, yielding a dumbbell-shaped complex of approximately 40 nm in length. Dissociation of the approximately 26S complexes releases in addition to approximately 20S cylinder particles a novel type of a disc-shaped particle of approximately 15 nm diameter which may represent the attached components or subcomplexes of them. Based on its structural and biochemical properties we postulate that the approximately 26S complex identified here is identical to the ubiquitin-dependent protease.
Peters
J M
JM
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg.
Harris
J R
JR
Kleinschmidt
J A
JA
D000818:Animals; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D003546:Cysteine Endopeptidases / Q000032:analysis / Q000302:isolation & purification / Q000648:ultrastructure*; D008854:Microscopy, Electron; D009097:Multienzyme Complexes / Q000032:analysis / Q000302:isolation & purification / Q000648:ultrastructure*; D016624:Negative Staining; D009865:Oocytes / Q000737:chemistry / Q000648:ultrastructure*; D046988:Proteasome Endopeptidase Complex; D014982:Xenopus laevis
D009097:Multienzyme Complexes; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex