Restricting α-Synuclein Transport into Mitochondria by Inhibition of α-Synuclein-VDAC Complexation as a Potential Therapeutic Target for Parkinson's Disease Treatment

Published

October 1, 2022

Author(s)

Megha Rajendran, Maria Queralt-Martin, Philip A. Gurnev, William M. Rosencrans, Amandine Rovini, Daniel Jacobs, Kaitlin Abrantes, David Hoogerheide, Sergey M. Bezrukov, Tatiana Rostovtseva

Abstract

Involvement of alpha-synuclein (αSyn) in Parkinson's disease (PD) is complicated and difficult to trace on cellular and molecular levels. Recently we established that αSyn can regulate mitochondrial function by voltage-activated complexation with the Voltage-Dependent Anion Channel (VDAC) of the outer mitochondrial membrane. When complexed with αSyn, the VDAC pore is partially blocked, reducing the transport of ATP/ADP and other metabolites. Further, αSyn can translocate into the mitochondria through VDAC, where it interferes with the mitochondrial respiration or function. Recruitment of αSyn to the VDAC-containing lipid membrane appears to be a crucial prerequisite for both the blockage and translocation processes. Here we report an inhibitory effect of HK2p, a small membrane-binding peptide from the mitochondria-targeting N-terminus of hexokinase 2, on the αSyn membrane binding, and hence on αSyn complex formation with VDAC and translocation through it. In electrophysiology experiments, addition of HK2p at micromolar concentrations to the same side of the membrane as αSyn results in dramatic reduction of the frequency of blockage events in a concentration-dependent manner, reporting on complexation inhibition. Using two complementary methods of measuring protein-membrane binding, bilayer overtone analysis and fluorescence correlation spectroscopy, we found that HK2p induces detachment of αSyn from lipid membranes. Experiments with live HeLa cells using proximity ligation assay confirmed that HK2p impedes αSyn entry into mitochondria. Our results demonstrate that it is possible to regulate αSyn-VDAC complexation by a rationally designed peptide, thus suggesting new avenues in the search for peptide therapeutics to alleviate αSyn mitochondrial toxicity in PD and other synucleinopathies.

Citation

Cellular and Molecular Life Sciences

Volume

79

Issue

10

Pub Type

Journals

Keywords

Membrane-binding peptide, synucleinopathies, hexokinase-2, voltage-dependent anion channel, single-channel electrophysiology, proximity ligation assay, small-peptide inhibitors

Citation

Rajendran, M. , Queralt-Martin, M. , Gurnev, P. , Rosencrans, W. , Rovini, A. , Jacobs, D. , Abrantes, K. , Hoogerheide, D. , Bezrukov, S. and Rostovtseva, T. (2022), Restricting α-Synuclein Transport into Mitochondria by Inhibition of α-Synuclein-VDAC Complexation as a Potential Therapeutic Target for Parkinson's Disease Treatment, Cellular and Molecular Life Sciences (Accessed April 23, 2024)