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Structural and Functional Interactions of Mitochondrial ADP-Phosphorylating Apparatus
Nůsková, Hana ; Houštěk, Josef (advisor) ; Kolarov, Jordan (referee) ; Kuda, Ondřej (referee) ; Panicucci Zíková, Alena (referee)
The complexes of the oxidative phosphorylation (OXPHOS) system in the inner mitochondrial membrane are organised into structural and functional super-assemblies, so-called supercomplexes. This type of organisation enables substrate channelling and hence improves the overall OXPHOS efficiency. ATP synthase associates into dimers and higher oligomers. Within the supercomplex of ATP synthasome, it interacts with ADP/ATP translocase (ANT), which exchanges synthesised ATP for cytosolic ADP, and inorganic phosphate carrier (PiC), which imports phosphate into the mitochondrial matrix. The existence of this supercomplex is generally accepted. Experimental evidence is however still lacking. In this thesis, structural interactions between ATP synthase, ANT and PiC were studied in detail. In addition, the interdependence of their expression was examined either under physiological conditions in rat tissues or using model cell lines with ATP synthase deficiencies of different origin. Specifically, they included mutations in the nuclear genes ATP5E and TMEM70 that code for subunit ε and the ancillary factor of ATP synthase biogenesis TMEM70, respectively, and a microdeletion at the interface of genes MT-ATP6 and MT-COX3 that impairs the mitochondrial translation of both subunit a of ATP synthase and subunit Cox3...
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Extraction of nucleotides from plant matrix and their determination by ion-exchange high performance liquid chromatography
Konečná, Markéta ; Sobotníková, Jana (advisor) ; Coufal, Pavel (referee)
In this thesis chromatographic conditions for the HPLC analysis of ribonucleotides AMP, ADP and ATP were optimized. For their separation chromatographic column, Nucleogel SAX 1000-8, 50 x 4,6 mm, from the German company Marcherey-Nagel was used. The mobile phase of a 0,2 mol/L solution of KH2PO4, its pH was adjusted with 1mol/L with potassium hydroxide to pH = 5,00. As the most suitable flow rate was selected 1,0 mL/min. The analysis was conducted using an isocratic elution. Detection was conducted by a DAD detector at a wavelength of λ = 260 nm. Substances were eluted in the following order AMP (tr = 0,911 min), ADP (tr = 1,667 min) and ATP (tr = 7,262 min). The total analysis time of the mixture of standards, under the conditions mentioned above, lasted for 10 minutes. For the extraction of adenosine-5-ribonucloetides the real matrix frozen and lyophilized leaves of tobacco virginia (Nicotiana tabacum L.) have been used. Two methods of extraction were applied. The first procedure was based on the extraction of AMP, ADP and ATP from the leaves of the tobacco plant by boiling deionized water. In the second method 0,07 mol/L HClO4 for the extraction instead of deionized water was used. Keywords: HPLC, adenosine ribonucleotides, AMP, ADP, ATP, tobacco virginia
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Structural and Functional Interactions of Mitochondrial ADP-Phosphorylating Apparatus
Nůsková, Hana ; Houštěk, Josef (advisor) ; Kolarov, Jordan (referee) ; Kuda, Ondřej (referee) ; Panicucci Zíková, Alena (referee)
The complexes of the oxidative phosphorylation (OXPHOS) system in the inner mitochondrial membrane are organised into structural and functional super-assemblies, so-called supercomplexes. This type of organisation enables substrate channelling and hence improves the overall OXPHOS efficiency. ATP synthase associates into dimers and higher oligomers. Within the supercomplex of ATP synthasome, it interacts with ADP/ATP translocase (ANT), which exchanges synthesised ATP for cytosolic ADP, and inorganic phosphate carrier (PiC), which imports phosphate into the mitochondrial matrix. The existence of this supercomplex is generally accepted. Experimental evidence is however still lacking. In this thesis, structural interactions between ATP synthase, ANT and PiC were studied in detail. In addition, the interdependence of their expression was examined either under physiological conditions in rat tissues or using model cell lines with ATP synthase deficiencies of different origin. Specifically, they included mutations in the nuclear genes ATP5E and TMEM70 that code for subunit ε and the ancillary factor of ATP synthase biogenesis TMEM70, respectively, and a microdeletion at the interface of genes MT-ATP6 and MT-COX3 that impairs the mitochondrial translation of both subunit a of ATP synthase and subunit Cox3...
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Digital Processing of Payroll Documents in accordance with German Legislation
Žižka, Jakub ; Kunstová, Renáta (advisor) ; Jakeš, Jiří (referee)
This thesis deals with archiving e-mail communication with attached accounting files in the environment of the German legislation at company ADP Employer Services. It also describes the laws which need to be taken into account and description of the product JobRouter. The aim of the thesis was on a detailed description of all processes related to digital archiving of payroll documents. Another objective was the description of key aspects of the German legislation, which must be taken into account in the system JobRouter. All these aspects had to be taken into account from the perspective from ADP and from the customer perspective. The work is divided into five parts. The first section describes the author's work for ADP and there is further elaborated the structure of work. In the second part is described in detail for ADP environment. The third part deals with all laws, standards and rules that need to be taken into account when the issue of payroll. In the fourth part the detailed analysis JobRouter are discussed in detail. The last part is the boot JobRouter evaluation and taken into account interaction with the law. The benefit of this work is describing the current state of archival documents in payroll for ADP. Also finding and describing all necessary laws to implement the new system JobRouter.
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