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National Repository of Grey Literature 1 records found  Search took 0.00 seconds. 
Preparation of human and schistosomal cathepsins L in enzymatically active form
 Horáková, Lenka ; Mareš, Michael (advisor) ; Ječmen, Tomáš (referee)
Cathepsin L-like proteases are involved in many pathological processes and their inhibitors are attractive molecules for the development of new drugs. This thesis focuses on human cathepsin L (hCL) and cathepsin L3 from the parasitic blood fluke Schistosoma mansoni (SmCL3). hCL is associated with cancer and Covid-19, while SmCL3 is the digestive enzyme of S. mansoni causing the disease schistosomiasis. The aim of the thesis was to prepare recombinant cathepsins L in an enzymatically active form that is sensitive to inhibitors. In the case of hCL, conditions for the controlled autoactivation of precursor forms to the mature form of the enzyme were identified and this pH- and temperature-dependent process was optimized. In the case of SmCL3, a protocol for purification of the mature form from the culture media of the yeast Komagataella pastoris was developed. The sensitivity of the prepared hCL and SmCL3 to inhibition was demonstrated using a model peptidomimetic vinyl sulfone whose IC50 values were in the sub/nanomolar concentration range. [IN CZECH] Key words: proteolytic enzymes, enzyme activity and inhibition, functional proteomics, recombinant protein expression, protein structure
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