The role of physiologically relevant metal ions on amyloid-beta peptide oligomerization and neurotoxicity in condition resembling amyloidosis (AMYLTOX)

 
 

AMYLTOX

This project is funded under the UEFISCDI research plan, The Development of the national system of research and development and is part of the “HUMAN RESOURCES” Program: Postdoctoral research project (PD).
Contract no. 25/2018 PN-III-P1-1.1-PD-2016-0737
The role of physiologically relevant metal ions on amyloid-beta peptide oligomerization and neurotoxicity in condition resembling amyloidosis.

Project Summary

Alzheimer’s disease (AD) belongs to a class of degenerative diseases commonly referred to as a protein misfolding diseases, whose main pathological features are progressive depositions of Aß peptides plaques between nerve cells, and neurofibrillary tangles within the nerve cells. Neurotoxic amyloid peptides are initially released as a monomers, and fibril formation starts with the formation of soluble Aß oligomer species. These small oligomeric amyloids were shown to allow ion permeability through ion channels-like structures inserted into the cellular membrane, and have proven to produce amyloid toxicity. Although, there is evidence for Aß oligomers being a “toxic species” and it is also clear that metals ions enhances amyloid peptide misfolding, it is not clear whether there is a physiological role for metal-biding or whether metal-binding occurs only during Aß formation. Furthermore, knowing that the properties of the membranes influence biological functions, the question rises whether the interaction between metal and lipid membranes modulate in some manner the Aß insertion and activity. Considering the above, this project proposes the following: (1) the study of the activity of Aß peptide oligomers in reconstituted lipid membranes, in condition resembling amyloidosis and (2) elucidate through which interfacial interactions between electrophysiological relevant metals and reconstituted lipid membranes alter the membrane affinity and activity of selected ß-amyloids.

Research Team

Project Director

Irina SCHIOPU, Scientific Researcher at the Alexandru Ioan Cuza University of Iasi, Interdisciplinary Research Institute, Laboratory of Molecular Biophysics And Medical Physics

Mentor

Lucel SIRGHI, Professor at the Scientific Researcher at the Alexandru Ioan Cuza University of Iasi, Physics Department, Leader of Iasi Plasma Advanced Research Center

Resources and Infrascture

Available Resources and Infrastructure

  • Complete electophysiology set-ups (Molecular Devices, HEKA); Patch-clamp amplifiers, Keithley electrometers
  • FluoroMax-4 spectrofluorimeter Horiba Jobin-Yvon
  • Axiovert 40 CFL inverted epi-fluorescence microscope
  • UV-VIS spectrophotometer
  • Peltier temperature regulator; PIP 5 pipette puller; ultraprecise motor micromanipulator; nanoliter injector; perfusion system for lipid bilayers
  • Liposomes manufacturing items (sonicator, chromatography columns, waveform generators, Avanti Mini-Extruder)
  • MPark SYSTEMS XE-100 Atomic Force Microscope with liquid cell with thermal control (ILX Lightwave LDT-5948 Precision Temperature Controller)
  • Dried vacuum pumps (Agilent Technologies SH-110 Dry Scroll Vacuum Pump) Turbomolecular vacuum pump; Vacuum components (pressure monitor VARIAN RGC-100, valves, flanges, etc)
  • Tektronix DPO2024B Digital Phosphor Oscilloscope
  • MBRAUN Glove Box GB 2202-P-D
  • Inverted confocal microscope Nikon Eclipse Ti-E equipped with TI-TIRF Illumination Unit, LU4 4 Laser Module with AOTF: 408 nm, 457-514 nm, 561nm; Andor iXon Camera CCD Du-860E; ACC-XW-CHIL-160 Oasis 160 Ultra compact chiller unit; GW Instek GFG-8250A Function Generator 5MHz
  • Parck Systems XE-100 70 or Nova NT-MDT Atomic Force Microscopy

Activities and Results

Published Papers and Conferences

Published papers

2020

 1.  Asandei Alina, Di Muccio Giovanni, Schiopu Irina, Mereuta Loredana, Dragomir Isabela, Chinappi Mauro, Luchian Tudor, Nanopore-Based Protein Sequencing Using Biopores: Current Achievements and Open Challenges, Small Methods, 2020, 1900595, pp. 1 – 13

2. Isabela Dragomir, Cezara Ioana Bucataru, Irina Schiopu, Tudor Luchian, Unzipping mechanism of free- and polyarginine-conjugated DNA-PNA duplexes, preconfined inside the a-hemolysin nanopore, Anal. Chem., 2020 (accepted)

2019
1. Loredana Mereuta, Alina Asandei, Irina Schiopu, Yoonkyung Park, Tudor Luchian, Nanopore-Assisted, Sequence-Specific Detection, and Single-Molecule Hybridization Analysis of Short, Single-Stranded DNAs, Anal. Chem., 2019, 91(13), pp. 8630-8637
2. Tudor Luchian, Yoonkyung Park, Alina Asandei, Irina Schiopu, Loredana Mereuta, Aurelia Apetrei, Nanoscale Probing of Informational Polymers with Nanopores. Applications to Amyloidogenic Fragments, Peptides, and DNA-PNA Hybrids, Acc. Chem. Res. 2019, 52(1), pp. 267-276

2018

Andrei Ciuca, Alina Asandei, Irina Schiopu, Aurelia Apetrei, Loredana Mereuta, Chang Ho Seo, Yoonkyung Park, and Tudor Luchian, Single-Molecule, Real-Time Dissecting of Peptide Nucleic Acid–DNA Duplexes with a Protein Nanopore Tweezer, Anal. Chem., 2018, 90 (12), pp. 7682–7690

Conferences

2019

International

1. Irina Schiopu, Ioana Cezara Bucataru, Tudor Luchian, Chang Ho Seo, Yoonkyung Park, Electrophoretic migration of PAMAM G1 dendrimers into confined nano-spaces: a single-molecule assay, 12th EBSA European Biophysics Congress / 10th IUPAP International Conference on Biological Physics (ICBP), 20-24 July 2019, Madrid, Spain (poster presentation)
2. Andrei Ciuca, Alina Asandei, Irina Schiopu, Loredana Mereuta, Chang Ho Seo, Yoonkyung Park and Tudor Luchian, Catch me, hug me, break me. Detecting and rupturing single DNA-PNA duplexes with a protein nanopore, Single-Molecule Sensors and NanoSystems International Conference – S3IC 2019, 03-05 April 2019, Munich, Germany (poster presentation)
3. Irina Schiopu, Ioana Cezara Bucataru, Tudor Luchian, Chang Ho Seo, Yoonkyung Park, Nanoscale investigation of low generation PAMAM dendrimers, 12TH INTERNATIONAL CONFERENCE Processes In Isotopes And Molecules and Satellite Workshop in the frame of PIM 2019, 24-27 September 2019, Cluj-Napoca, Romania (poster presentation)
4. Loredana Mereuta, Alina Asandei, Irina Schiopu, Yoonkyung Park, Tudor Luchian, Hybridization-Based Detection and Validation of ssDNA fragments, with a Protein Nanopore, Single Molecule Protein Sequencing 2019, 16-19 September 2019, Jerusalim, Israel (poster presentation)

2018

National

Irina Schiopu, Alina Asandei, Andrei Ciuca, Aurelia Apetrei, Loredana Mereuta, Chang-Ho Seo, Yoonkyung Park and Tudor Luchian, Study of pH Mediated Nanoscale Confinement of Small Generation Dendrimers with Protein Nanopores, 15th National Conference of Biophysics with international participation, September 7 – 10, 2018, Bucharest, Romania (poster presentation)

Andrei Ciuca, Alina Asandei, Irina Schiopu, Aurelia Apetrei, Loredana Mereuta, Chang-Ho Sea, Yoonkyung Park and Tudor Luchian, Breaking Apart Molecular Dimers with Nanopores, 15th National Conference of Biophysics with international participation, September 7 – 10, 2018, Bucharest, Romania (poster presentation)

Prizes

2018

Best Poster Prize at the 15th National Conference of Biophysics with international participation, September 7 – 10, 2018, Bucharest, Romania, for the work: Breaking Apart Molecular Dimers with Nanopores

Contact

Address

“Alexandru Ioan Cuza” University of Iași
Department For Research And Project Management
Carol I no. 11 Boulevard
700506, Iași, Romania

Contact

Phone: +40 (232) 201 024 / +40 (232) 201 023
Fax: +40 (232) 201 201

Interdisciplinary Research Institute
Scientific Researcher III Irina ȘCHIOPU, Ph.D.
Phone: +40 (232) 201 191
E-mail: irina.schiopu@mail.uaic.ro