|
|
Formation of carbonyl compounds by non-enzymatic protein glycation
Krautová, Lenka ; Boušová, Iva (advisor) ; Dršata, Jaroslav (referee)
Author: Lenka Krautová Publishing place: Charles University in Prague, Faculty of Pharmacy in Hradec Králové Title: Formation of carbonyl compounds by non-enzymatic protein glycation Supervisor: PharmDr. Iva Boušová, Ph.D. This diploma thesis was aimed at the optimization of carbonyl groups determination in biological material. Carbonyl groups are formed by effect of protein oxidation or glycation. As a model protein, I used bovine serum albumin for spectrophotometric and aspartate aminotransferase for electrophoretic method of assessment, which were glycated by several glycating agents (glucose; glucose + copper; fructose; fructose + copper; methylglyoxal). I optimized different parameters of spectrophotometric determination: solvent, concentration of reagent, amount of protein, and duration of incubation. I managed to find suitable conditions of this determination, which I have used in subsequent experiments with different glycating agents. At electrophoretic determination, I performed SDS-PAGE with following silver staining or immunoblotting with chemiluminescent detection. The results of spectrophotometric determination showed that methylglyoxal has the highest glycating ability, followed by fructose with addition of cupric ions, fructose itself, glucose with addition of cupric ions, and the...
|
|
| |
|
| |
|
|
Formation of carbonyl compounds by non-enzymatic protein glycation
Krautová, Lenka ; Boušová, Iva (advisor) ; Dršata, Jaroslav (referee)
Author: Lenka Krautová Publishing place: Charles University in Prague, Faculty of Pharmacy in Hradec Králové Title: Formation of carbonyl compounds by non-enzymatic protein glycation Supervisor: PharmDr. Iva Boušová, Ph.D. This diploma thesis was aimed at the optimization of carbonyl groups determination in biological material. Carbonyl groups are formed by effect of protein oxidation or glycation. As a model protein, I used bovine serum albumin for spectrophotometric and aspartate aminotransferase for electrophoretic method of assessment, which were glycated by several glycating agents (glucose; glucose + copper; fructose; fructose + copper; methylglyoxal). I optimized different parameters of spectrophotometric determination: solvent, concentration of reagent, amount of protein, and duration of incubation. I managed to find suitable conditions of this determination, which I have used in subsequent experiments with different glycating agents. At electrophoretic determination, I performed SDS-PAGE with following silver staining or immunoblotting with chemiluminescent detection. The results of spectrophotometric determination showed that methylglyoxal has the highest glycating ability, followed by fructose with addition of cupric ions, fructose itself, glucose with addition of cupric ions, and the...
|
guest ::
