UittrekselCalpains have drawn much attention because of their crucial role in many biological processes as well as in many diseases caused by abnormal calpain activity. Calpains are proteases that are regulated by calcium and calpastatin, an endogenous calpain inhibitor. Among calpain family members, m-calpain is ubiquitously expressed in mammalian cells and is the focus of many studies. Calpastatin is an intrinsically disordered protein (IDP), which is a new important class of proteins not only because of their flexibility in interacting with various partners but alsobecause of their critical role in biological processes. The number of newly discovered IDPs is still increasing and they are very new in the field of protein design
The aim of this thesis is to analyze the inhibitory effect of the in silico designed artificial IDPs against calpain activity. For this purpose, human m-calpain and human calpastatin domain1 (hCSD1) were recombinantly produced and characterized. 2 artificial IDPs, which are expected to inhibit calpain activity as a designed property were purified. In addition, 27 artificial peptides, which were computationally designed to have the capacity of binding to the surface of m-calpain with calpain. Therefore, no inhibition of calpain activity by the artificial designed IDPs could be observed. From the data that was generated during this thesis, it was concluded that the artificial designed constructs need to be reevaluated for the aim of understanding why there was no interaction between the artificial IDPs with calpain nor inhibition of the calpain activity by the artificial IDPs.
|Datum Prijs||6 sep 2013|
|Begeleider||Peter Tompa (Promotor) & Kris Pauwels (Co-promotor)|