Inhibition of amyloid beta-induced synaptotoxicity by a pentapeptide derived from the glycine zipper region of the neurotoxic peptide

doi:10.1016/j.neurobiolaging.2013.06.001

Neurobiology of Aging

Volume 34, Issue 12, December 2013, Pages 2805-2814

https://doi.org/10.1016/j.neurobiolaging.2013.06.001 Get rights and content

Abstract

A major characteristic of Alzheimer's disease is the presence of amyloid beta (Aβ) oligomers and aggregates in the brain. Aβ oligomers interact with the neuronal membrane inducing perforations, causing an influx of calcium ions and increasing the release of synaptic vesicles that leads to a delayed synaptic failure by vesicle depletion. Here, we identified a neuroprotective pentapeptide anti-Aβ compound having the sequence of the glycine zipper region of the C-terminal of Aβ (G₃₃LMVG₃₇). Docking and Förster resonance energy transfer experiments showed that G₃₃LMVG₃₇ interacts with Aβ at the C-terminal region, which is important for Aβ association and insertion into the lipid membrane. Furthermore, this pentapeptide interfered with Aβ aggregation, association, and perforation of the plasma membrane. The synaptotoxicity induced by Aβ after acute and chronic applications were abolished by G₃₃LMVG₃₇. These results provide a novel rationale for drug development against Alzheimer's disease.

Introduction

Alzheimer's disease is a progressive, neurodegenerative pathology, which affects the central nervous system in the elderly, causing cognitive and behavioral disorders. A major histopathological feature of this disease is the presence of macroscopic structures in the brain parenchyma called senile plaques, which consist mainly of oligomers of amyloid beta (Aβ) (Kawahara et al., 2000). These oligomers can directly alter the plasma membrane through the formation of a pore-like structure that allows the passage of calcium ions (Arispe et al., 1993, Sepulveda et al., 2010). The mechanism of this membrane insertion is largely unknown, but the early calcium entry increases the release of synaptic vesicles in hippocampal neurons, thus facilitating neurotransmission (Parodi et al., 2009). Chronic application of Aβ, in contrast, reduces the number of synaptic vesicles leading to neurotransmission failure (Parodi et al., 2009).

The Aβ protein sequence contains amino acids with properties like metal coordination (Nakamura et al., 2007), aggregation (Armstrong et al., 2011), and membrane insertion (Kim, 2005) (Fig. 1A). It is widely believed that the hydrophobic C-terminal of Aβ interacts with the lipid bilayer and the N-terminal contains polar, charged residues facing the solvated pore formed by Aβ oligomers (Jang et al., 2008). Though most of the small peptides used to block Aβ toxicity were focused on the N-terminal sequence of Aβ, the C-terminal region has been largely ignored. The physiopathological effect of Aβ pores was demonstrated using NA7 (EVHHQKL), a minipeptide derived from the N-terminus of Aβ, that blocked calcium and cell toxicity mediated by Aβ (Arispe et al., 2008, Sepulveda et al., 2010). The C-terminal region, in contrast, is important for association and insertion of Aβ into the lipid membrane, and G33 and G37 have been described as important in the process of Aβ association with artificial lipid membranes (Hung et al., 2008). Furthermore, the pathogenic effects of Aβ have been attributed to the hydrophobic C-terminal stretch of 14 amino acids that is also thought to be the seed for aggregation (Jarrett and Lansbury, 1993). Within this region, 3 repeat GxxxG motifs encompassing Aβ residues G25–G37 (G25xxxG29xxxG33xxxG37) promote Aβ aggregation independent of the hydrophobic residues in the x positions (Kim and Hecht, 2006). Interestingly, these glycines might be important in the formation of ion-permeable Aβ channels (Kim, 2005) and they can promote dimerization of the amyloid precursor protein (Barnham et al., 2006, Kienlen-Campard et al., 2008, Munter et al., 2007).

The aim of the present study was to develop a new anti-Aβ compound based on a pentapeptide having the sequence G₃₃LMVG₃₇ and that was able to block the aggregation and the synaptoxicity induced by Aβ. In addition, Förster resonance energy transfer (FRET) and docking studies with monomer (Crescenzi et al., 2002), fibril (Luhrs et al., 2005), and the crystallographically resolved structure of a partial sequence of Aβ (Streltsov et al., 2011), support the idea of a direct interaction of Aβ with the G₃₃LMVG₃₇ peptide. The inhibition of these pathophysiological processes by a small peptide, similar to those used in cancer, endocrinological diseases, and neurological disorders (Bidwell, 2012, Dodel et al., 2010, Herrero et al., 2012), could be beneficial in altering the course of AD.

Section snippets

Peptide preparation and storage

Human Aβ_1–42 labeled with 6-carboxyfluorescein (FAM; green fluorescence) or 6-carboxytetramethylrhodamine (TAMRA; red fluorescence) at its N-terminus, and unlabeled peptides were purchased from Anaspec. Aβ_1–42 was dissolved in hexafluoroisopropanol (HFIP, 10 mg/mL) or dimethyl sulfoxide (DMSO, 10 mg/mL) and stored in aliquots at −20 °C. For the preparation of Aβ aggregates (80-μM), aliquots of 5 μL were added to 137.5 μL ultrapure water in an Eppendorf tube. After 10–20-minute incubation at

Interaction of GLMVG with Aβ

We performed molecular docking assays to predict some structural components involved in the interaction between G₃₃LMVG₃₇ and Aβ. For these studies, we used structures of G₃₃LMVG₃₇ (red) and Aβ (yellow, protein data bank:1IYT) (Fig. 1B) (Crescenzi et al., 2002). The analysis of G₃₃LMVG₃₇-Aβ complexes suggests that G₃₃LMVG₃₇ interacts with the C-terminal of the Aβ structure (Fig. 1B). Detailed analysis showed that G₃₃LMVG₃₇ associated with Aβ in different conformations, that can be summarized in

Discussion

Although there exists some debate on whether Aβ can up- or downregulate selective components of synaptic transmission, several studies point out that Aβ alters pre- and postsynaptic components affecting long-term potentiation, N-methyl-D-aspartate (NMDA) and 2-amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl) propanoic acid (AMPA) neurotransmissions and calcium homeostasis (Glabe and Kayed, 2006, Parodi et al., 2009, Stephan et al., 2001). Supporting the complexity of these effects, no single

Disclosure statement

The authors declare that they have no conflicts of interest. Animal use protocols were approved by the University of Concepcion Bioethics Committee (Approval #2004100A).

Acknowledgements

The authors thank Lauren Aguayo for revising the manuscript and for technical assistance, and Dr José Martínez for assistance in FRET experiments. This work was supported by FONDECYT grant 1100502 (LGA, CO). C.P. and C.F.B. are CONICYT (Comision Nacional de Investigacion Cientifica y Tecnologica) fellows.

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¹: Present address: Oxidation Biology Laboratory, The Florey Institute of Neuroscience and Mental Health, The University of Melbourne, Parkville, Victoria, Australia.

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Regular articleInhibition of amyloid beta-induced synaptotoxicity by a pentapeptide derived from the glycine zipper region of the neurotoxic peptide

Abstract

Introduction

Section snippets

Peptide preparation and storage

Interaction of GLMVG with Aβ

Discussion

Disclosure statement

Acknowledgements

Biophys. J.

J. Biol. Chem.

Trends Biochem. Sci.

Cell

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Neurosci. Lett.

J. Biol. Chem.

J. Biol. Chem.

J. Mol. Biol.

Gramicidin channels

IEEE Trans. Nanobioscience

Alzheimer disease amyloid beta protein forms calcium channels in bilayer membranes: blockade by tromethamine and aluminum

Proc. Natl. Acad. Sci. U.S.A

Mutations that replace aromatic side chains promote aggregation of the Alzheimer's Abeta peptide

Biochemistry

Prefibrillar amyloid aggregates could be generic toxins in higher organisms

J. Neurosci.

Peptides for cancer therapy: a drug-development opportunity and a drug-delivery challenge

Ther. Deliv.

Anti-homeostatic synaptic plasticity of glycine receptor function after chronic strychnine in developing cultured mouse spinal neurons

J. Neurochem.

ZDOCK: an initial-stage protein-docking algorithm

Proteins

ClusPro: a fully automated algorithm for protein-protein docking

Nucleic Acids Res.

Solution structure of the Alzheimer amyloid beta-peptide (1-42) in an apolar microenvironment. Similarity with a virus fusion domain

Eur. J. Biochem.

Annular alpha-synuclein protofibrils are produced when spherical protofibrils are incubated in solution or bound to brain-derived membranes

Biochemistry

Intravenous immunoglobulins as a treatment for Alzheimer's disease: rationale and current evidence

Drugs

Peptides as drugs

QJM

Common structure and toxic function of amyloid oligomers implies a common mechanism of pathogenesis

Neurology

Regular article
Inhibition of amyloid beta-induced synaptotoxicity by a pentapeptide derived from the glycine zipper region of the neurotoxic peptide