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Structure determination of helical domain of DNA damage-inducible protein 2
Staníček, Jakub ; Grantz Šašková, Klára (advisor) ; Obšil, Tomáš (referee)
Human Ddi2 and his homologues are scarcely explored family of ubiquitin- like/ubiquitin-associated domain proteins (UBL/UBA proteins), containing domain which is structurally related to dimeric aspartyl proteases from retroviruses. The most studied of this family is Ddi1 protein from Saccharomyces cerevisiae, which functions within the ubiquitin- proteasome system. This key cellular system exploits tightly regulated enzymatic apparatus for highly selective posttranslational modifications of proteins with molecules of ubiquitin, which serves as intracellular signal determining the proteins fate, importantly its degradation in many cases. Ddi1 plays a role of proteasome adaptor within this context, helping the recognition of ubiquitylated proteins by the proteasome. Even though the function as a proteasome receptor is possible for human Ddi2 protein as well, its cellular role might have been altered during the evolution. One of the important steps in decoding its purpose in cell is exploration of function of its individual domains. This work focuses on structural study of neighbouring region of this protease domain, where the helix-rich region called HDD domain is located. This region of Ddi2 was cloned, expressed and subjected to the NMR measurements. Structural information based on the NMR data was...
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Searching for a physiological partner of Ddi2 (DNA damage-inducible protein homolog 2) protein
Kurfürst, Jaroslav ; Grantz Šašková, Klára (advisor) ; Vaněk, Ondřej (referee)
One of the most important cellular processes, essential not only for protein degradation, is the so called ubiquitin-proteasome system. A key player in this system is ubiquitin, a small protein with unique ability to form various chains. Cellular proteins marked for degradation via ubiquitin, are recruited to the proteasome either by a direct interaction with one of the intrinsic proteasomal receptors, or by adaptor proteins. These proteins typically possess ubiquitin-like domain and ubiquitin associated domain that predispose them for delivering ubiquinated proteins to the proteasome. Adaptor protein called Ddi2 differs from other members of this family by possessing additional domain called the retroviral protease like domain. This domain is structurally similar to HIV protease and its proteolytic function has been discovered only recently. Due to the presence of this proteolytic domain one could expect that Ddi2 might be a deubiquitinase. Here we therefore tested the possible cleavage of diubiquitin chains by recombinantly prepared Ddi2 protein. We can conclude that Ddi2 did not show any deubiquitinating activity in given conditions.
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Structure determination of helical domain of DNA damage-inducible protein 2
Staníček, Jakub ; Grantz Šašková, Klára (advisor) ; Obšil, Tomáš (referee)
Human Ddi2 and his homologues are scarcely explored family of ubiquitin- like/ubiquitin-associated domain proteins (UBL/UBA proteins), containing domain which is structurally related to dimeric aspartyl proteases from retroviruses. The most studied of this family is Ddi1 protein from Saccharomyces cerevisiae, which functions within the ubiquitin- proteasome system. This key cellular system exploits tightly regulated enzymatic apparatus for highly selective posttranslational modifications of proteins with molecules of ubiquitin, which serves as intracellular signal determining the proteins fate, importantly its degradation in many cases. Ddi1 plays a role of proteasome adaptor within this context, helping the recognition of ubiquitylated proteins by the proteasome. Even though the function as a proteasome receptor is possible for human Ddi2 protein as well, its cellular role might have been altered during the evolution. One of the important steps in decoding its purpose in cell is exploration of function of its individual domains. This work focuses on structural study of neighbouring region of this protease domain, where the helix-rich region called HDD domain is located. This region of Ddi2 was cloned, expressed and subjected to the NMR measurements. Structural information based on the NMR data was...
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