|
| |
|
|
Regulation of the activity of aspartic and serine proteases by selective natural inhibitors
Srp, Jaroslav ; Mareš, Michael (advisor) ; Novák, Petr (referee) ; Bařinka, Cyril (referee)
Proteases are involved in many physiological processes and their dysregulation is associated with various pathologies. Protease activity is effectively controlled by natural inhibitors. This PhD thesis is focused on the inhibitors of aspartic and serine proteases of animal and plant origin and provides the identification, biochemical characterization and structural description of their inhibition mechanisms. Plant Kunitz inhibitors are produced as defensive proteins, and they are able to block activities of a broad spectrum of proteases. In this thesis, the digestive proteolytic system of the Colorado potato beetle, a herbivore pest of potato plants, was described with the help of functional proteomics. It was shown that aspartic and serine proteases from this herbivore are effectively blocked by two potato Kunitz inhibitors (namely PCDI, PSPI). Using structural analysis, novel types of reactive centers were identified on PCDI and PSPI molecules for the inhibition of aspartic protease cathepsin D and the serine proteases trypsin and chymotrypsin. The analysis of the reactive center on a PCDI with the crystal structure of digestive cathepsin D from the Colorado potato beetle explained the mechanism of their interaction. Sphingolipids were identified as the first endogenous inhibitors of human...
|
|
|
ENUM client
Srp, Jaroslav ; Peterka, Jiří (advisor) ; Bednárek, David (referee)
For node signing and addressing in the internet the 32 bits number is used - known as IP address. Because the long numbers can be hardly memorize the abstract known as domain name is established. This abstraction transforms the domain name to the IP address via DNS (the reverse process is known as reverse DNS). But not everybody owns fixed awarded IP address, that is a problem. This situation is not true for the phone numbers (fixed or mobile phones), of course. The ENUM system which is introduced in this graduation theses uses the reverse DNS to save the information (web, e-mail, sip and other addresses, phone numbers for MMS routing and such like records) to the set phone or other number. After it you can send a query to the reverse DNS for the set phone number and by return you get the mentioned informations about the phone number. As addendum are implemented the applications (client and server) and web browser plug-in that can be used for ENUM system testing, using and exploiting.
|
|
|
Regulation of the activity of aspartic and serine proteases by selective natural inhibitors
Srp, Jaroslav
Proteases are involved in many physiological processes and their dysregulation is associated with various pathologies. Protease activity is effectively controlled by natural inhibitors. This PhD thesis is focused on the inhibitors of aspartic and serine proteases of animal and plant origin and provides the identification, biochemical characterization and structural description of their inhibition mechanisms. Plant Kunitz inhibitors are produced as defensive proteins, and they are able to block activities of a broad spectrum of proteases. In this thesis, the digestive proteolytic system of the Colorado potato beetle, a herbivore pest of potato plants, was described with the help of functional proteomics. It was shown that aspartic and serine proteases from this herbivore are effectively blocked by two potato Kunitz inhibitors (namely PCDI, PSPI). Using structural analysis, novel types of reactive centers were identified on PCDI and PSPI molecules for the inhibition of aspartic protease cathepsin D and the serine proteases trypsin and chymotrypsin. The analysis of the reactive center on a PCDI with the crystal structure of digestive cathepsin D from the Colorado potato beetle explained the mechanism of their interaction. Sphingolipids were identified as the first endogenous inhibitors of human...
|
|
|
Regulation of the activity of aspartic and serine proteases by selective natural inhibitors
Srp, Jaroslav
Proteases are involved in many physiological processes and their dysregulation is associated with various pathologies. Protease activity is effectively controlled by natural inhibitors. This PhD thesis is focused on the inhibitors of aspartic and serine proteases of animal and plant origin and provides the identification, biochemical characterization and structural description of their inhibition mechanisms. Plant Kunitz inhibitors are produced as defensive proteins, and they are able to block activities of a broad spectrum of proteases. In this thesis, the digestive proteolytic system of the Colorado potato beetle, a herbivore pest of potato plants, was described with the help of functional proteomics. It was shown that aspartic and serine proteases from this herbivore are effectively blocked by two potato Kunitz inhibitors (namely PCDI, PSPI). Using structural analysis, novel types of reactive centers were identified on PCDI and PSPI molecules for the inhibition of aspartic protease cathepsin D and the serine proteases trypsin and chymotrypsin. The analysis of the reactive center on a PCDI with the crystal structure of digestive cathepsin D from the Colorado potato beetle explained the mechanism of their interaction. Sphingolipids were identified as the first endogenous inhibitors of human...
|
|
|
Regulation of the activity of aspartic and serine proteases by selective natural inhibitors
Srp, Jaroslav ; Mareš, Michael (advisor) ; Novák, Petr (referee) ; Bařinka, Cyril (referee)
Proteases are involved in many physiological processes and their dysregulation is associated with various pathologies. Protease activity is effectively controlled by natural inhibitors. This PhD thesis is focused on the inhibitors of aspartic and serine proteases of animal and plant origin and provides the identification, biochemical characterization and structural description of their inhibition mechanisms. Plant Kunitz inhibitors are produced as defensive proteins, and they are able to block activities of a broad spectrum of proteases. In this thesis, the digestive proteolytic system of the Colorado potato beetle, a herbivore pest of potato plants, was described with the help of functional proteomics. It was shown that aspartic and serine proteases from this herbivore are effectively blocked by two potato Kunitz inhibitors (namely PCDI, PSPI). Using structural analysis, novel types of reactive centers were identified on PCDI and PSPI molecules for the inhibition of aspartic protease cathepsin D and the serine proteases trypsin and chymotrypsin. The analysis of the reactive center on a PCDI with the crystal structure of digestive cathepsin D from the Colorado potato beetle explained the mechanism of their interaction. Sphingolipids were identified as the first endogenous inhibitors of human...
|
|
| |
|
|
ENUM client
Srp, Jaroslav ; Bednárek, David (referee) ; Peterka, Jiří (advisor)
For node signing and addressing in the internet the 32 bits number is used - known as IP address. Because the long numbers can be hardly memorize the abstract known as domain name is established. This abstraction transforms the domain name to the IP address via DNS (the reverse process is known as reverse DNS). But not everybody owns fixed awarded IP address, that is a problem. This situation is not true for the phone numbers (fixed or mobile phones), of course. The ENUM system which is introduced in this graduation theses uses the reverse DNS to save the information (web, e-mail, sip and other addresses, phone numbers for MMS routing and such like records) to the set phone or other number. After it you can send a query to the reverse DNS for the set phone number and by return you get the mentioned informations about the phone number. As addendum are implemented the applications (client and server) and web browser plug-in that can be used for ENUM system testing, using and exploiting.
|
|
| |
|
|
Digestive aspartic protease of Colorado beetle
Srp, Jaroslav ; Jonáková, Věra (referee) ; Konvalinka, Jan (advisor)
Colorado potato beetle (Leptinotarsa decemlineata) is an economically important herbivorous pest. Cathepsin D-like aspartic peptidase (LdCD) plays an important role during protein degradation in the midgut of Colorado potato beetle. This work describes the preparation of two expression systems, namely in Escherichia coli and Pichia pastoris, for the production of recombinant LdCD. The protocol for refolding of denatured LdCD was designed and optimized. Activation of the inactive LdCD zymogen and cleavage of the propetide (activation peptide) were investigated. This process proceeds autocatalytically at acidic pH or with the assistance of the cysteine peptidase legumain. The proteolytic activity of LdCD was characterized using fluorogenic peptidic substrate and protein substrates, and kinetic parameters and pH optimum were determined. The inhibition specificity of LdCD was analyzed using a panel of peptidase inhibitors. LdCD was significantly inhibited by PDI (potato cathepsin D inhibitor), a protein inhibitor produced in potato leaves. This suggests that PDI is a natural defense protein, which is directed against LdCD in the midgut of Colorado potato beetle in order to block the digestion. The potential application of PDI in the construction of transgenic crops resistant against insects is discussed.
|
guest ::
RSS feed.