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Carbonylation of human serum albumin by glycation agents: effect of carbonyl reductase 1
Mazurková, Veronika ; Boušová, Iva (advisor) ; Skálová, Lenka (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Veronika Mazurková Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of diploma thesis: Carbonylation of human serum albumin by glycation agents: effect of carbonyl reductase 1 This diploma thesis is concerned with a formation of carbonyl groups in the molecules of human serum albumin (HSA) as a result of selected glycation agents activity and the ability of enzyme carbonyl reductase 1 (CBR1) to influence this process. Initial experiments were devoted to find optimal conditions for quantification of carbonyl groups in the model of HSA glycated in vitro by fructose and methylglyoxal using spectrophotometric assays and the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with subsequent immunoblotting. The glycation of HSA was monitored also by the measurement of fluorescent AGE products. Then the effect of CBR1 on the formation of carbonyl groups in the samples of HSA glycated with reactive carbonyl compounds fructose, methylglyoxal, glyoxal, and glyceraldehyde was studied. Spectrophotometric assays, SDS-PAGE with immunoblotting and enzyme immunoassay ELISA were employed. CBR1 was not able to remove already formed carbonyl groups under given conditions. Also the...
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Effect of selected sesquiterpenes on carbonyl reductase 1 activity in vitro
Mazurková, Veronika ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Veronika Mazurková Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of rigorous thesis: Effect of selected sesquiterpenes on carbonyl reductase 1 activity in vitro Sesquiterpenes, the main components of plant essential oils, are integral parts of spices, traditional food and beverages. They are used in folk medicines, pharmaceutical industry as well as in cosmetics. Several sesquiterpenes possess interesting biological activities but they could interact with concurrently administered drugs via the modulation of activity and/or expression of drug-metabolizing enzymes. Enzyme carbonyl reductase 1 (CBR1) is one of the enzymes participating in the metabolism of a number of endogenous and xenobiotic compounds. One of the CBR1 substrates is chemotherapeutic agent doxorubicin, which is metabolized by this enzyme to the less effective metabolite doxorubicinol, which is responsible for the doxorubicin cardiotoxicity. The aim of this rigorous thesis was to find out the ability of five selected sesquiterpenes (i.e. valencene, β-caryophyllene oxide, α-humulene, cis-nerolidol and trans-nerolidol) to affect the activity of CBR1 in vitro. The effect of studied substances on the activity of this enzyme was...
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Carbonylation of human serum albumin by glycation agents: effect of carbonyl reductase 1
Mazurková, Veronika ; Boušová, Iva (advisor) ; Skálová, Lenka (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Veronika Mazurková Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of diploma thesis: Carbonylation of human serum albumin by glycation agents: effect of carbonyl reductase 1 This diploma thesis is concerned with a formation of carbonyl groups in the molecules of human serum albumin (HSA) as a result of selected glycation agents activity and the ability of enzyme carbonyl reductase 1 (CBR1) to influence this process. Initial experiments were devoted to find optimal conditions for quantification of carbonyl groups in the model of HSA glycated in vitro by fructose and methylglyoxal using spectrophotometric assays and the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with subsequent immunoblotting. The glycation of HSA was monitored also by the measurement of fluorescent AGE products. Then the effect of CBR1 on the formation of carbonyl groups in the samples of HSA glycated with reactive carbonyl compounds fructose, methylglyoxal, glyoxal, and glyceraldehyde was studied. Spectrophotometric assays, SDS-PAGE with immunoblotting and enzyme immunoassay ELISA were employed. CBR1 was not able to remove already formed carbonyl groups under given conditions. Also the...
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