Abstract (eng)
As the number of cases of Alzheimer’s disease (AD) is constantly rising in all developed countries, the unmet medical need for a curative treatment continues to grow. Current drugs improve symptoms, but do not have profound disease-modifying effects. The generation and aggregation of β-amyloid peptides (Aβ), proteolytic derivatives of amyloid precursor protein (APP), is believed to play a key role in the onset and progression of AD. Consequently, inhibition of the major APP cleaving enzyme, BACE1 (β-site APP cleaving enzyme), is generally recognised as a promising therapeutic strategy and currently an active area of investigation. A wide variety of BACE1 inhibitors have been reported so far. Among them is Reticulon 3 (RTN3), which was recently proposed to act as a natural cellular modulator of BACE1 activity. This study focused particularly on the interaction between BACE1 and RTN3, aiming to reconstitute the original approach and confirm binding between the two proteins. When using a transiently transfected neuroblastoma cell line binding assays did not show the proposed interaction. In a different approach, representing a fundamental part of the project, RTN3 was successfully cloned and expressed recombinantly. A systematic method was devised to combine both protein purification and binding studies of RTN3 and BACE1. Surprisingly, my findings indicated that the proposed interaction cannot be reconstituted using recombinant proteins.