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Cloning, expression and characterization of human serine racemase mutants
Nováková, Ilona ; Brynda, Jiří (referee) ; Konvalinka, Jan (advisor)
AAbbssttrraacctt Human serine racemase (hSR) is a cytosolic pyridoxal-5'-phosphate dependent enzyme localized in the central nervous system. It synthesizes D-serine, which is an endogenous coagonist for the N-methyl-D-aspartate (NMDA) receptors and plays a key role in excitatory neurotransmission in the brain. Thus, human serine racemase is a promising target for the treatment of neurodegenerative diseases connected with NMDA receptors. However, few specific inhibitors have been identified to date and the crystal structure of hSR has become available only very recently. We decided to perform a random mutagenesis to determine the amino acid residues critical for the enzyme activity. Ser 84 was reported as a catalytic residue along with Lys 56. After analysis of a double mutant S84G/P111L which retained its capability to convert L-serine to pyruvate, we prepared and characterized the single mutant S84G in order to exclude potential effect of the P111L mutation.on the activity of the analyzed enzyme. KKeeyy wwoorrddss:: D-serine; Serine racemase; PLP-dependent enzymes; Random mutagenesis; Racemases
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Inhibition of Thymidine Phosphorylase
Zákoucká, Eva ; Brynda, Jiří (advisor) ; Obšil, Tomáš (referee)
2. Abstract Thymidine phosphorylase (TPase), also known as gliostatin or Platelet-derived endothelial cell growth factor (PD-ECGF), is an enzyme with an important role in the nucleoside metabolism and is also involved in degradation and recycling of DNA. TPase catalyzes the reversible phosphorolysis of pyrimidine 2'-deoxynucleosides to 2-deoxy-D-ribose-1- phosphate and their respective bases, as well as the transfer of the deoxyribosyl moiety from one pyrimidine base to another. Thymidine phosphorylase is a therapeutic target of great importance because of its participation in angiogenesis especially in solid tumors of various tissues. Therefore, TPase stimulates tumor growth and progression, as well as metastasis. In addition to this, TPase inhibits apoptosis, particularly of tumor cells and causes degradation of several antiviral and anticancer drugs. Apart from the carcinoma tissues, thymidine phosphorylase is overexpressed in various other tissues affected by disorders characterized by proliferation of blood vessels including psoriasis, rheumatoid arthritis and atherosclerosis. Inhibiting the activity of TPase selectively in the tissues affected by the diseases listed above would be of great therapeutic significance. Therefore, many inhibitors, mainly substrate analogues, have been designed based on the...
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Production of mouse recombinant prion protein and analysis of its properties
Krejčiříková, Anna ; Brynda, Jiří (referee) ; Konvalinka, Jan (advisor)
Title: Production of mouse recombinant prion protein and analysis of its properties Author: Anna Krejčiříková Department: Department of Biochemistry Supervisor: doc. RNDr. Jan Konvalinka, CSc., Supervisor's e-mail: konval@uochb.cas.cz Consultant: dr. Ing. Karel Holada, IIM 1stFM CU Abstract: Although prion diseases represent only a small fraction of all known neurodegenra- tive illnesses, they deserve our attention mainly due to the fact that they are lethal, incurable by today, and having a potential to cause a serious epidemic. Even though this topic has been studied for many years, there are still many unanswered questions concerning both the mechanism, and the spread of prion diseases. The most promising current theory is the protein-only hypothesis, which explains the fundamental of the ill- ness by the conversion of cellular prion protein (PrPC ) into pathological prion protein (PrPSc ). The important tool in proving this theory is also a recombinant prion pro- tein. The presented thesis summarizes existing successes in the proving of protein-only hypothesis. In the experimental part, we prepared the mouse recombinant prion pro- tein (mrPrP) in E. coli bacteria. The structure of the purified and renaturated protein was verified by CD spectroscopy. Next, we focus on the effects of the...
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Thermodynamic Limits for Dehalogenation of Producer Gas by Na2CO3, NaAlO2, CaO/CaCO3, BaO/BaCO3 and Mixed CaO-BaO/BaCO3 Sorbents.
Svoboda, Karel ; Leitner, J. ; Havlica, Jaromír ; Brynda, Jiří ; Skoblia, S. ; Hartman, Miloslav ; Pohořelý, Michael ; Hsu, H.-T.
Our study is focused on thermodynamic analyses and minimization of Gibbs energy for gas de-halogenation (removal of HCl, HF and HBr) by selected solid sorbents (Na2CO3, NaAlO2, CaO/CaCO3, BaO/BaCO3, and mixtures containing CaO-BaO/BaCO3).
Fulltext: content.csg -  PDF
Plný tet: SKMBT_C22016090211393 - PDF
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Multi-stage Biomass Gasification
Skoblia, S. ; Beňo, Z. ; Brynda, J. ; Pohořelý, Michael ; Picek, I.
The presented work describes the specific design and operational characteristics of the multi-stage (two-stage) gasifiers developed by Tarpo ltd. The article contains the operation parameters, typical gas and tar composition, for different operation conditions, shows the effect of power output and properties of wood chips to gas quality.
Fulltext: content.csg - PDF
Plný tet: SKMBT_C22015111911040 - PDF
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