Opis projektu
Atak z udziałem białka opiekuńczego na agregaty białkowe
Składanie się białek ma kluczowe znaczenie dla ich prawidłowego funkcjonowania, a w procesie tym pomagają inne białka zwane opiekuńczymi. Coraz więcej danych wskazuje na to, że białka opiekuńcze rozbijają ponadto agregaty białkowe, jakie tworzą się w trakcie procesu neurodegeneracji w przebiegu chorób Alzheimera, Parkinsona i pląsawicy Huntingtona. Jednak mechanizm rozbijania agregatów białkowych pozostaje w dużej mierze nieznany. Celem badaczy z finansowanego ze środków UE projektu NMR-DisAgg jest zbadanie mechanizmów interakcji białek opiekuńczych i pokonanie ograniczeń związanych z dynamiką i krótkotrwałym charakterem tych oddziaływań. Opisanie reakcji rozbijania agregatów da lepszy wgląd w proces degeneracji i być może przyczyni się do zidentyfikowania nowych celów terapeutycznych.
Cel
Molecular chaperones are a diverse group of proteins critical to maintaining cellular homeostasis. Aside from protein refolding, it has recently been discovered that certain combinations of human chaperones can break apart toxic protein aggregates and even amyloids that have been linked to a host of neurodegenerative diseases. The first chaperones in this disaggregation reaction that are responsible for recognizing and performing initial remodeling of aggregates, are members of the Hsp40 (DnaJ) and small heat shock protein (sHSP) families. Very little, though, is known regarding how these chaperones perform their functions, and characterization of sHsp- and DnaJ-substrate complexes by most structural techniques has proven extremely challenging, as most chaperones are dynamic in nature and typically operate through a series of transient interactions with both their clients and other chaperones.
The advanced NMR techniques used in our lab, however, are ideally suited for the study of these exact types of dynamic systems, and include recently developed experiments (CEST, CPMG) that allow us to monitor the transient and low populated protein states typical of chaperone-chaperone and chaperone-client interactions, as well as to study the structure of these potentially very large protein complexes (methyl-TROSY).
By exploiting these NMR methodologies and additional, novel labeling schemes, we will characterize, for the first time, the recognition and substrate remodeling performed by the many members of the DnaJ and sHsp chaperone families on their clients. We will then take these approaches one step further and develop real time NMR experiments to observe the client remodeling performed over the course of the disaggregation reaction itself.
By combining advanced NMR with biophysical and functional assays, we ultimately aim to identify the specific sets of chaperones that, with the Hsp70 system, protect our cells by dissolving disease-linked aggregates and amyloid fibers.
Dziedzina nauki
- engineering and technologymaterials engineeringfibers
- natural scienceschemical sciencesinorganic chemistrytransition metals
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesbiological sciencescell biology
- natural sciencesphysical sciencesopticsspectroscopyabsorption spectroscopy
Słowa kluczowe
Program(-y)
Temat(-y)
System finansowania
ERC-STG - Starting GrantInstytucja przyjmująca
7610001 Rehovot
Izrael