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Schlagwörter:
Alzheimer Disease/etiology/metabolism Amino Acid Motifs Animals Cells Cells, Cultured Dendritic Spines/metabolism/pathology Humans Mice Microscopy, Atomic Force Mutant Proteins/chemistry/genetics/metabolism Neurons/*metabolism/*pathology Phosphorylation Protein Conformation Protein Multimerization Protein Structure, Tertiary Repetitive Sequences, Amino Acid tau Proteins/*chemistry/genetics/*metabolism
Zusammenfassung:
Several neurodegenerative diseases are characterized by the aggregation and posttranslational modifications of Tau protein. Its "repeat domain" (TauRD) is mainly responsible for the aggregation properties, and oligomeric forms are thought to dominate the toxic effects of Tau. Here we investigated the conformational transitions of this domain during oligomerization and aggregation in different states of beta-propensity and pseudo-phosphorylation, using several complementary imaging and spectroscopic methods. Although the repeat domain generally aggregates more readily than full-length Tau, its aggregation was greatly slowed down by phosphorylation or pseudo-phosphorylation at the KXGS motifs, concomitant with an extended phase of oligomerization. Analogous effects were observed with pro-aggregant variants of TauRD. Oligomers became most evident in the case of the pro-aggregant mutant TauRDDeltaK280, as monitored by atomic force microscopy, and the fluorescence lifetime of Alexa-labeled Tau (time-correlated single photon counting (TCSPC)), consistent with its pronounced toxicity in mouse models. In cell models or primary neurons, neither oligomers nor fibrils of TauRD or TauRDDeltaK280 had a toxic effect, as seen by assays with lactate dehydrogenase and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, respectively. However, oligomers of pro-aggregant TauRDDeltaK280 specifically caused a loss of spine density in differentiated neurons, indicating a locally restricted impairment of function.
