Keeping up the PICUP
(2019) KEMT30 20191Department of Chemistry
- Abstract
- The aggregation of α-synuclein into fibrils has been long proven to be closely related to the toxicity leading to Parkinson’s disease. In this process, medium-sized intermediates called oligomers are formed, a heterogeneous group of aggregates formed by a small number of monomers. While fibrils have been proven to cause little to no toxicity, oligomers have been demonstrated to induce cell death. This has pointed at oligomers as the possible key in understanding the relationship between protein and disease. However, its concentration during aggregation process in comparison to monomers and fibrils has made its analysis a difficult task. Photo induced crosslinking of unmodified proteins (PICUP) is a method that has been previously used for... (More)
- The aggregation of α-synuclein into fibrils has been long proven to be closely related to the toxicity leading to Parkinson’s disease. In this process, medium-sized intermediates called oligomers are formed, a heterogeneous group of aggregates formed by a small number of monomers. While fibrils have been proven to cause little to no toxicity, oligomers have been demonstrated to induce cell death. This has pointed at oligomers as the possible key in understanding the relationship between protein and disease. However, its concentration during aggregation process in comparison to monomers and fibrils has made its analysis a difficult task. Photo induced crosslinking of unmodified proteins (PICUP) is a method that has been previously used for oligomer analysis. By crosslinking oligomers together, their stability is ensured, and they can be subjected to methods that couldn’t be used otherwise, making it possible to perform population analysis on. However, the method doesn’t have a unified protocol nor a set instrument in the market as of yet. The main goal of this research was to optimize the PICUP method by creating a more reproducible instrument of easy access to other research groups, as well as optimizing conditions such as reagents or light exposure time. Finally, the reaction was used to evaluate the oligomer population distribution throughout the aggregation of α-synuclein. The results proved the newly created instrument and the optimized conditions to be optimal for oligomer analysis, opening the gate to a big range of future experiments which will help us unravel the mysteries behind the aggregation process and its connection with cellular toxicity. (Less)
- Popular Abstract
- Proteins are one of the most essential components of life. From controlling division or transporting “cargo” inside a cell, to coordinating the movement of a multicellular organism, proteins are responsible for most of the “active” parts of life. In order to keep us working, cells constantly produce big amounts of the specific protein we need in each part of our body. However, some of those protein’s functions have yet to be completely understood. Sometimes a protein is found because of its connection to a disease before we learn what it actually does. That’s the case for α-synuclein, a small protein produced in the area called substantia nigra in our brain. This protein was first discovered in some inclusion bodies called Lewy bodies,... (More)
- Proteins are one of the most essential components of life. From controlling division or transporting “cargo” inside a cell, to coordinating the movement of a multicellular organism, proteins are responsible for most of the “active” parts of life. In order to keep us working, cells constantly produce big amounts of the specific protein we need in each part of our body. However, some of those protein’s functions have yet to be completely understood. Sometimes a protein is found because of its connection to a disease before we learn what it actually does. That’s the case for α-synuclein, a small protein produced in the area called substantia nigra in our brain. This protein was first discovered in some inclusion bodies called Lewy bodies, which could be found on Parkinson’s patients. Ever since then, the relationship between protein and disease has been thoroughly investigated.
One of the biggest discoveries in that field has been the fact that the protein aggregating into big fibril structures has been linked to its toxicity. In that aggregation process, the protein units (monomers) cluster together forming many different sized aggregates. Oligomers, medium-sized aggregates consisting of 2 to around 20 units of proteins, have recently been proven to cause cell death. However, its concentration when compared to pure monomers or the big fibrils is very low. Due to this, many techniques have failed to analyze oligomers. Photo induced crosslinking of unmodified proteins (PICUP) is a very promising method which can be used for oligomer analysis. In this reaction, when light is shone on a protein-containing sample, units forming the oligomer bind together tightly, allowing us to perform methods to analyze them which we couldn’t otherwise use. However, this method hasn’t been standardized and optimized, to the point of there not even being an available instrument in the market to perform the technique.
The main goal of this project was to develop an instrument to perform PICUP, and then proceed to optimize conditions of the reaction, such as testing different reagents or different durations for the light to shine on the reaction. Finally, when these conditions were optimized, the technique was used to analyze how the population of oligomers changes throughout the α-synuclein aggregation process. This didn’t only prove the instrument and the optimization to be a very useful tool in analyzing oligomers, but it threw more light into an important aspect of aggregation. All in all, the project has proven PICUP to be a very useful technique to help solve the many questions we have yet to answer in order to understand the connection between the protein and Parkinson’s disease. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/8982480
- author
- Ortigosa, Lei LU
- supervisor
- organization
- alternative title
- The paparazzi of aggregating proteins
- course
- KEMT30 20191
- year
- 2019
- type
- H1 - Master's Degree (One Year)
- subject
- keywords
- α-synuclein Parkinson's disease Crosslinking, protein science, proteinvetenskap
- language
- English
- id
- 8982480
- date added to LUP
- 2019-07-04 13:33:09
- date last changed
- 2019-07-04 13:33:09
@misc{8982480,
abstract = {{The aggregation of α-synuclein into fibrils has been long proven to be closely related to the toxicity leading to Parkinson’s disease. In this process, medium-sized intermediates called oligomers are formed, a heterogeneous group of aggregates formed by a small number of monomers. While fibrils have been proven to cause little to no toxicity, oligomers have been demonstrated to induce cell death. This has pointed at oligomers as the possible key in understanding the relationship between protein and disease. However, its concentration during aggregation process in comparison to monomers and fibrils has made its analysis a difficult task. Photo induced crosslinking of unmodified proteins (PICUP) is a method that has been previously used for oligomer analysis. By crosslinking oligomers together, their stability is ensured, and they can be subjected to methods that couldn’t be used otherwise, making it possible to perform population analysis on. However, the method doesn’t have a unified protocol nor a set instrument in the market as of yet. The main goal of this research was to optimize the PICUP method by creating a more reproducible instrument of easy access to other research groups, as well as optimizing conditions such as reagents or light exposure time. Finally, the reaction was used to evaluate the oligomer population distribution throughout the aggregation of α-synuclein. The results proved the newly created instrument and the optimized conditions to be optimal for oligomer analysis, opening the gate to a big range of future experiments which will help us unravel the mysteries behind the aggregation process and its connection with cellular toxicity.}},
author = {{Ortigosa, Lei}},
language = {{eng}},
note = {{Student Paper}},
title = {{Keeping up the PICUP}},
year = {{2019}},
}