ausblenden:
Schlagwörter:
CRYOELECTRON TOMOGRAPHY; ANTIGEN PRESENTATION; GIANT PROTEASE; PHASE
PLATE; DEGRADATION; CLASSIFICATION; VISUALIZATION; SUBTOMOGRAMS;
DROSOPHILA; TOOLBOXScience & Technology - Other Topics; primary cultured neuronal cell; 26S proteasome; Volta phase plate;
cryo-electron tomography; subtomogram averaging;
Zusammenfassung:
Tripeptidyl peptidase II (TPPII) is a eukaryotic protease acting downstream of the 26S proteasome; it removes tripeptides from the degradation products released by the proteasome. Structural studies in vitro have revealed the basic architecture of TPPII, a two-stranded linear polymer that assembles to form a spindle-shaped complex of similar to 6 MDa. Dependent on protein concentration, TPPII has a distinct tendency for polymorphism. Therefore, its structure in vivo has remained unclear. To resolve this issue, we have scrutinized cryo-electron tomograms of rat hippocampal neurons for the occurrence and spatial distribution of TPPII by template matching. The quality of the tomograms recorded with the Volta phase plate enabled a detailed structural analysis of TPPII despite its low abundance. Two different assembly states (36-mers and 32-mers) coexist as well as occasional extended forms with longer strands. A distance analysis of the relative locations of TPPII and 26S proteasomes confirmed the visual impression that these two complexes spatially associate in agreement with TPPII's role in postproteasomal degradation.
