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Exploration of extraterrestrial body impact and UV-driven transformation of atmospheres of terrestrial planets
Knížek, Antonín ; Ferus, Martin (advisor) ; Kabáth, Petr (referee) ; Kopecký, Vladimír (referee)
Keywords: Exoplanets, Earth, Impact, Photochemistry, Infrared Spectroscopy, Atmospheric chemistry Impacts and photochemistry are two very important driving forces for chemical transformation of planetary atmospheres. While strong UV radiation produced by young stars continuously provides a significant amount of energy, impacts are one-time events with far-reaching consequences. Especially important are then impacts on young rocky planets, including the early Earth, because these planets are likely exposed to much higher impact fluxes, commonly called 'heavy bombardment'. This bombardment is the final echo of the turbulent planetary accretion and has prominent significance for planetary environments, e.g., the chemical composition and shape of the planetary surface, the chemistry of atmospheres, aerosol production, and likely the origin of life. Future observations of exoplanets by space telescopes, such as the James Webb Space Telescope or Ariel, as well as ground-based telescopes, such as the Extremely Large Telescope currently under construction in Chile, could determine whether this heavy bombardment represents a common scenario for the evolution of terrestrial planets. Both impacts and photochemistry can be efficiently simulated and studied in laboratory conditions by state-of-the-art methods....
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Experimental study of chemical evolution of biomolecules under early Earth conditions
Knížek, Antonín ; Ferus, Martin (advisor) ; Adam, Vojtěch (referee)
Origin of life is a still-enduring gap in human knowledge. This work is focused on revealing of several pieces of this puzzle. State of the art scenarios of biomolecules synthesis under prebiotic conditions are presented and discussed. This thesis presents our recent experiments suggesting a novel idea that neutral planetary atmosphere containing a mixture of neutral volcanic-type gasses (CO2, N2, H2O) can be converted over acidic mineral catalysts upon irradiation by a soft UV-radiation into a relatively reactive mixture of reducing gases (CH4, CO), which can be further reprocessed by high-energy chemistry. The resulting mixture (CH4, CO + N2) represents a common reducing atmosphere related e.g. to the chemistry of Titan, the largest moon of Saturn, as well as a possible representation of the secondary atmosphere of our planet. Also, photocatalytic reduction of CO2-rich atmosphere can explain the abiotic origin of methane on current Mars or other terrestrial planets. In our subsequent experiments, corresponding equimolar model mixture of CH4 : CO : N2 in presence of water vapour was subjected to reprocessing by high-power laser plasma simulating an asteroid impact - one of a series of impact events which the young Earth experienced during the first 600 million years of her history. Upon delivery...
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Expression and purification of protein photo-initiated nanoprobe: tool to study clinically relevant protein-protein interactions
Knížek, Antonín ; Šulc, Miroslav (advisor) ; Koblihová, Jitka (referee)
Cytochrome b5 is a key protein in the function and regulation of the mixed function monooxygenase (MFO) system in mammalian endoplasmic reticulum and is, therefore, a clinically relevant target for biochemical studies. To study its interactions within the MFO system using photo-initiated crosslinking, we have developed cytochrome b5 mutants with methionine in several key amino acid positions within the primary amino acid sequence, such as serine 23 and leucine 41. Also, naturally presented Met in positions 96, 126 and 131 were mutated to Leu with no effect to cytochrome b5 activity. Our protein was expressed in E. coli B834 auxotrophic type with L-2-amino-5,5-azi-hexanoic acid (photo-Met) present in the cultivation medium. This methionine analogue with photolabile diazirine ring is readily incorporated in Met positions into the primary sequence of proteins by aminoacyl-tRNA synthetases. The whole expression protocol was optimized to achieve maximal percentage of photo-Met incorporation into the expressed protein sequence. Up to 93.4% incorporation of photo-Met was achieved. The expressed protein was isolated and photo-Met incorporation was established with MALDI-TOF mass spectrometry. After reconstitution with its natural interaction partners - full-length cytochrome P450 2B4 (rabbit isoform) or...
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