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Vinsamlegast notið þetta auðkenni þegar þið vitnið til verksins eða tengið í það: https://hdl.handle.net/1946/23814

Titill: 
  • Titill er á ensku Synthesis of L68Q mutant cystatin C. Aggregation and autophagy in HEK293T cells
Námsstig: 
  • Meistara
Útdráttur: 
  • Útdráttur er á ensku

    Hereditary cystatin C amyloid angiopathy (HCCAA) is an incurable dominant hereditary disease that affects the arteries of the brain and causes recurrent strokes in carriers. The average lifespan of carriers is about 30 years. The disease is classified as one of the cerebral amyloid angiopathies, which are characterized by deposition of protein amyloid. In the case of HCCAA there is deposition of the cystatin C protein, due to mutation L68Q, in the arterial walls of the brain. Finding a cure for these diseases is of importance since their prevalence increases gradually from the age of 65, mostly due to Alzheimer’s disease. In this study the aim was to find ways to produce both human wild type and L68Q mutant cystatin C. The main source of the latter, especially in the form of amyloid, has been post mortem extraction from brain tissue of patients. A comparison was made of several physical properties that might play a role in disease pathogenesis, i.e. aggregation tendency and effects on autophagy. The wild type and L68Q mutant variant of cystatin C were produced in HEK293T cells. The proteins were subsequently isolated from media by filtration through beaded cellulose columns. The results show that the mutant variant has an increased tendency to aggregate and is more rapidly degraded in conditioned media from HEK293T cells. These results are in compliance with previously published results. Both variants seem to induce autophagy in HEK293T cells but the mutant possibly to a lesser extent. It can be concluded from these results that HEK293T cells are suitable for the production of secreted human proteins for research. Furthermore, it can be concluded that the pathology of HCCAA can possibly be linked to different physical properties of L68Q cystatin C found in patients with HCCAA and normal cystatin C protein.

Samþykkt: 
  • 19.2.2016
URI: 
  • http://hdl.handle.net/1946/23814


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