Characterization of the molecular mechanism behind CTL - the major choline uptake system in eukaryotes

Informacje na temat projektu

CTL

Identyfikator umowy o grant: 890822

DOI

10.3030/890822

Projekt został zamknięty

Data podpisania przez KE 23 Marca 2020

Data rozpoczęcia 1 Maja 2020

Data zakończenia 30 Kwietnia 2022

Finansowanie w ramach

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Koszt całkowity

€ 219 312,00

Wkład UE

€ 219 312,00

Koordynowany przez

AARHUS UNIVERSITET
Denmark

Periodic Reporting for period 1 - CTL (Characterization of the molecular mechanism behind CTL - the major choline uptake system in eukaryotes)

Okres sprawozdawczy: 2020-05-01 do 2022-04-30

Podsumowanie kontekstu i ogólnych celów projektu

Choline is an essential nutrient for the human body with a critical importance for the fetal development of the brain, the nervous system and metabolism. It is a precursor in the synthesis of membrane phospholipids, the neurotransmitter acetylcholine and the methyl donor betaine. In order to be available for the human body it needs to be transported through the plasma membrane by specialized proteins, called membrane transporters. Transmembrane uptake of choline is the essential, but still enigmatic, first step in choline metabolism in the cell. Furthermore, uptake and cellular homeostasis of choline is affected in several diseases (e.g. Alzheimer, cardiovascular disease, fetal alcohol syndrome), brain ischemic events, aging and cancer; underlining its importance for human health.
The choline transporter-like (CTL) family forms a unique class of membrane transporters with five family members, that facilitate the majority of choline uptake in all eukaryotes. With no direct homologies to other transporter families, protein structures, especially with bound substrate, are of utmost importance to understand how this enigmatic transporter class facilitates substrate transport. Therefore, the overall objectives of this project were the structural, functional and biochemical characterization of CTL proteins.

Prace wykonane od początku projektu do końca okresu sprawozdawczego oraz najważniejsze dotychczasowe rezultaty

During the funding period I worked extensively on proteins of the CTL family, towards their structural and functional characterization. I was able to establish the recombinant expression of several CTL proteins. I solubilized CTL proteins using different detergents from their membrane environment and purified these using chromatography methods (e.g. affinity chromatography, size exclusion chromatography).
Further sample analysis was conducted using biochemical and structural methods. Uptake assays for instance, were performed using xenopus oocytes expressing CTL proteins. Radioactive compounds were presented to the oocytes and uptake was measured by quantifying the radioactivity that was accumulated inside the oocytes.

Innowacyjność oraz oczekiwany potencjalny wpływ (w tym dotychczasowe znaczenie społeczno-gospodarcze i szersze implikacje społeczne projektu)

We have not published our results yet, but we are currently finalizing the first paper on our finding on CTL proteins and we expect to publish another paper, that will deepen our knowledge on the choline transporter-like family.

Choline uptake by CTL proteins