2002
Application of capillary zone electrophoresis for the study and assay of enzymes
NOVÁKOVÁ, Soňa a Zdeněk GLATZZákladní údaje
Originální název
Application of capillary zone electrophoresis for the study and assay of enzymes
Autoři
Vydání
2002. vyd. Brno, Book of abstracts of 6th Meeting of biochemists and molecular biologists, s. 77-77, 2002
Nakladatel
MU Brno
Další údaje
Jazyk
angličtina
Typ výsledku
Stať ve sborníku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Česká republika
Utajení
není předmětem státního či obchodního tajemství
Organizační jednotka
Přírodovědecká fakulta
Klíčová slova anglicky
EMMA; enzymes
Změněno: 19. 5. 2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.
Anotace
V originále
Ten years ago a new application for the evaluation of enzymatic reactions in capillary electrophoresis was proposed and developed, electrophoretically mediated microanalysis (EMMA). In this method, substrate(s) and enzyme are introduced in the capillary as distinct plugs, the first analyte injected being the one with the lower electrophoretic mobility. Upon the application of an electric field, these two zones interpenetrate due the differences in their electrophoretic mobilities. Enzymatic reaction takes place and the resultant reaction product(s) and the unreacted substrate(s) are electrophoretically transported towards detector, where they are individually detected. Since its discovery by Bao and Regnier the EMMA methodology has been utilized in a number of biochemical systems: for assays of enzyme activities, determinations substrates, Michaelis constants, inhibitors and inhibition constants, etc. In this communication the combination of the EMMA methodology with a partial filling technique was applied to study the kinetic parameters of bi-substrate enzymatic reaction of rhodanese. In this set-up the part of the capillary is filled with the buffer best for the enzymatic reaction whereas the rest of the capillary with the background electrolyte optimal for separation of substrates and products. The basic limitation of EMMA methodology the necessity to have the electrophoretic conditions compatible with both the separation of substrate(s) and product(s) of the enzymatic reaction and the enzymatic reaction is thus overcome.
Návaznosti
GA525/00/0785, projekt VaV |
|