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Modulation of the ankyrin TRPA1 receptor by divalent cations
Hynková, Anna ; Hudeček, Jiří (advisor) ; Hodek, Petr (referee)
TRPA1 is the only mammalian member of the ankyrin type subfamily of transient receptor potential (TRP) ion channels. This channel is widely expressed in many cell types and tissues, including sensory neurones, heart, brain, and lung, where it detects stimuli of different sensory modalities, including a broad spectrum of potentially harmful irritants and pungent compounds, such as isothiocyanates, thiosulfinates, acrolein, cinnamaldehyde derivatives, eugenol, nicotine, as well as multiple products of oxidative stress. In addition to these chemicals, TRPA1 channel can be also activated and, concurrently, inhibited by intracellular calcium, however, the exact mechanisms of this regulation are still unclear. TRPA1 is a nonselective cation channel, permeable for calcium ions, however, relative scattered data on TRPA1 channel selectivity provide divergent estimates ranging from 0,84 to 7,9; in addition, these values strongly depend on the activation mode of the channel. It appears that the extent to which calcium ions dynamically modulate TRPA1 depends on the total amount of cations permeating the ion channel pore. This diploma thesis contributes to the understanding of this topic by (i) investigating the effects of extracellular calcium ions on chemically and voltage- induced activity of human TRPA1...
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Binding of organic dyes to proteins. Applications in practical course in biochemistry.
Hynková, Anna ; Hudeček, Jiří (advisor) ; Koblihová, Jitka (referee)
In this submitted thesis the possibility of creating a new laboratory task for advanced practical courses in biochemistry, concerning the binding of a low-molecular compound (dye) to a protein macromolecule, was experimentally verified. First intention was to modify the existing task "Dialysis kinetics" with a binding of fluorescein to a protein. However, the experiments have proved, that fluorescence measurements of this kind are not reproducible in the conditions of practical courses laboratory and absorbance measurements have low sensitivity. Therefor a whole new task was created: "Binding of bromophenol blue to serum albumin", in which the stoichiometry of this binding is studied using so-called Job plot. After the optimization of procedure a laboratory task instructions were created, which are attached to this thesis. Keywords: fluorescein, bromophenol blue, serum albumin
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Modulation of the ankyrin TRPA1 receptor by divalent cations
Hynková, Anna ; Hudeček, Jiří (advisor) ; Hodek, Petr (referee)
TRPA1 is the only mammalian member of the ankyrin type subfamily of transient receptor potential (TRP) ion channels. This channel is widely expressed in many cell types and tissues, including sensory neurones, heart, brain, and lung, where it detects stimuli of different sensory modalities, including a broad spectrum of potentially harmful irritants and pungent compounds, such as isothiocyanates, thiosulfinates, acrolein, cinnamaldehyde derivatives, eugenol, nicotine, as well as multiple products of oxidative stress. In addition to these chemicals, TRPA1 channel can be also activated and, concurrently, inhibited by intracellular calcium, however, the exact mechanisms of this regulation are still unclear. TRPA1 is a nonselective cation channel, permeable for calcium ions, however, relative scattered data on TRPA1 channel selectivity provide divergent estimates ranging from 0,84 to 7,9; in addition, these values strongly depend on the activation mode of the channel. It appears that the extent to which calcium ions dynamically modulate TRPA1 depends on the total amount of cations permeating the ion channel pore. This diploma thesis contributes to the understanding of this topic by (i) investigating the effects of extracellular calcium ions on chemically and voltage- induced activity of human TRPA1...
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Binding of organic dyes to proteins. Applications in practical course in biochemistry.
Hynková, Anna ; Hudeček, Jiří (advisor) ; Koblihová, Jitka (referee)
In this submitted thesis the possibility of creating a new laboratory task for advanced practical courses in biochemistry, concerning the binding of a low-molecular compound (dye) to a protein macromolecule, was experimentally verified. First intention was to modify the existing task "Dialysis kinetics" with a binding of fluorescein to a protein. However, the experiments have proved, that fluorescence measurements of this kind are not reproducible in the conditions of practical courses laboratory and absorbance measurements have low sensitivity. Therefor a whole new task was created: "Binding of bromophenol blue to serum albumin", in which the stoichiometry of this binding is studied using so-called Job plot. After the optimization of procedure a laboratory task instructions were created, which are attached to this thesis. Keywords: fluorescein, bromophenol blue, serum albumin
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