Descrizione del progetto
Recettori sintetici per il trasporto di nucleotidi attraverso le membrane biologiche
Le cellule sono circondate da membrane biologiche che fungono da barriere rispetto all’ambiente circostante, ma favoriscono anche la comunicazione, consentendo ad esempio il trasporto selettivo di molecole. Il trasporto ionico nelle cellule non avviene passivamente, ma è effettuato da proteine vettrici di ioni dedicate che attraversano la membrana cellulare. L’obiettivo principale del progetto ORGANITRA, finanziato dall’UE, è sviluppare recettori sintetici in grado di trasportare composti di fosfati e fosforilati, come i nucleotidi, attraverso le membrane. Utilizzando substrati e chimica combinatoria, i ricercatori genereranno recettori sintetici che potranno essere utilizzati per introdurre selettivamente i nucleotidi nei liposomi e nelle cellule. I risultati di ORGANITRA apriranno nuove possibilità nei campi della biochimica e della biotecnologia.
Obiettivo
This ORGANITRA project addresses transmembrane transport. Lipid bilayer membranes not only define the borders of cells and their compartments but are also implicated in metabolic processes and signal transduction. Membranes function as impermeable barriers for ionic and hydrophilic species which can only cross the membrane with the aid of dedicated membrane proteins.
For biotechnological and biophysical applications, the development of anion carriers that can bind an anion and transport it across the lipid bilayer could be of great relevance. In this project, synthetic anion receptors will be developed to bind biologically relevant organic phosphorylated compounds, like nucleotides. These receptors will then be used to transport these organophosphates across membranes.
The receptors will be synthesised by dynamic combinatorial chemistry. Building blocks containing urea or thiourea groups, for efficient phosphate binding, will be connected to multi-armed scaffolds by hydrazone groups. The dynamic character of these bonds will be used to identify efficient receptors from libraries of compounds, using different phosphorylated compounds as templates. With this approach, selective receptors for different nucleotides and related compounds can be obtained.
The transport performance of the receptors will be evaluated with newly developed assays. Liposomes will be used as model systems and transport will be monitored by fluorescence spectroscopy using the quenching of the emission of an encapsulated phosphate sensitive dye. Additionally, the mechanism of the transport processes will be elucidated by fluorescence and 1H and 31P NMR spectroscopies.
Transmembrane carriers for phosphorylated compounds will make it possible to selectively introduce nucleotides into liposomes and cells, opening the way to fuel enzymes with adenosine triphosphate (ATP) in liposomes as biotechnological nanoreactors and to study nucleotide-dependent biochemical processes in cells.
Campo scientifico
- natural sciencesphysical sciencesopticsspectroscopyemission spectroscopy
- natural sciencesbiological sciencesbiochemistrybiomoleculeslipids
- natural sciencesphysical sciencesopticsspectroscopyabsorption spectroscopy
- natural sciencesbiological sciencesgeneticsnucleotides
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes
Programma(i)
Argomento(i)
Meccanismo di finanziamento
ERC-STG - Starting GrantIstituzione ospitante
1050 Bruxelles / Brussel
Belgio