Multicolor, Cell-Impermeable, and High Affinity BACE1 Inhibitor Probes Enable Superior Endogenous Staining and Imaging of Single Molecules

Stockinger F, Poc P, Möhwald A, Karch S, Häfner S, Alzheimer C, Sandoz G, Huth T, Broichhagen J (2024)

Publication Type: Journal article

Publication year: 2024

Journal

Journal of Medicinal Chemistry American Chemical Society

DOI: 10.1021/acs.jmedchem.4c00339

Abstract

The prevailing but not undisputed amyloid cascade hypothesis places the β-site of APP cleaving enzyme 1 (BACE1) center stage in Alzheimer′s Disease pathogenesis. Here, we investigated functional properties of BACE1 with novel tag- and antibody-free labeling tools, which are conjugates of the BACE1-inhibitor IV (also referred to as C3) linked to different impermeable Alexa Fluor dyes. We show that these fluorescent small molecules bind specifically to BACE1, with a 1:1 labeling stoichiometry at their orthosteric site. This is a crucial property especially for single-molecule and super-resolution microscopy approaches, allowing characterization of the dyes′ labeling capabilities in overexpressing cell systems and in native neuronal tissue. With multiple colors at hand, we evaluated BACE1-multimerization by Förster resonance energy transfer (FRET) acceptor-photobleaching and single-particle imaging of native BACE1. In summary, our novel fluorescent inhibitors, termed Alexa-C3, offer unprecedented insights into protein-protein interactions and diffusion behavior of BACE1 down to the single molecule level.

Involved external institutions

University of Côte d'Azur / Université Côte d'Azur FR France (FR) Max-Planck-Institut für medizinische Forschung / Max Planck Institute for Medical Research DE Germany (DE)

How to cite

APA:

Stockinger, F., Poc, P., Möhwald, A., Karch, S., Häfner, S., Alzheimer, C.,... Broichhagen, J. (2024). Multicolor, Cell-Impermeable, and High Affinity BACE1 Inhibitor Probes Enable Superior Endogenous Staining and Imaging of Single Molecules. Journal of Medicinal Chemistry. https://doi.org/10.1021/acs.jmedchem.4c00339

MLA:

Stockinger, Florian, et al. "Multicolor, Cell-Impermeable, and High Affinity BACE1 Inhibitor Probes Enable Superior Endogenous Staining and Imaging of Single Molecules." Journal of Medicinal Chemistry (2024).

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