D 2003

Application of Capillary Zone Electrophoresis for the Study of Enzymes

GLATZ, Zdeněk, Soňa NOVÁKOVÁ a Magdalena TELNAROVÁ

Základní údaje

Originální název

Application of Capillary Zone Electrophoresis for the Study of Enzymes

Autoři

GLATZ, Zdeněk (203 Česká republika, garant), Soňa NOVÁKOVÁ (203 Česká republika) a Magdalena TELNAROVÁ (203 Česká republika)

Vydání

Nice, France, Proceeding of 27th Symposium on HPLC and Related Techniques, od s. 133-133, 1 s. 2003

Nakladatel

MCI Paris France

Další údaje

Jazyk

angličtina

Typ výsledku

Stať ve sborníku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Francie

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/03:00008124

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

CZE; Enzymes

Štítky

Změněno: 19. 5. 2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.

Anotace

V originále

Enzymes are biological catalysts that play an important role in biochemical reactions necessary for normal growth, maturation and reproduction through whole live world. Due to their low concentrations in the samples containing large amount of other proteins, direct measurements of enzymes by masses are impossible. However enzymes can be measured more easily by their catalytic activities, which are the most relevant properties of enzymes in the biochemical context. The accurate measurement of enzymatic activity in biological samples is important in many fields of biochemistry, not only in routine biochemistry and in fundamental research but also in clinical and pharmacological research and diagnosis. The enzyme assay is also important procedure in elucidation of enzyme properties and function. Determination of kinetic parameters is usually undertaken to characterize an enzyme, to provide a quantitative evaluation of substrate specificity and to study kinetic mechanisms. In the last decade capillary electromigration separation techniques have become powerful techniques, which can provide highly efficient separations and large peak capacities. Today the availability of sophisticated automated equipment for CZE makes the technique suitable to be implement in routine laboratories for separation of a variety of molecules including metal ions, organic and inorganic ions, drugs, peptides, proteins and DNA fragments. All these parameters predict CZE as one of the method of choice also for determination of enzyme activities. Measurements of enzyme activity by CZE, which have been under active investigation in recent years, include the enzyme reaction prior CZE analysis and the enzyme reaction during CZE also known as electrophoretically mediated microanalysis (EMMA). Both these approaches were applied in the complex study on rhodanese, important enzyme catalysing detoxication of cyanide to less toxic thiocyanate after reaction with a sulfur donor, such as thiosulfate. The enzyme activity of rhodanese and the effects of temperature and pH on enzymatic reaction were evaluated by the off-capillary approach, whereas the kinetic parameters such as the Michaelis and inhibition constants were determined by means of the EMMA methodology.

Návaznosti

GA203/03/1125, projekt VaV
Název: Využití kapilární zónové elektroforézy pro studium enzymů
Investor: Grantová agentura ČR, Využití kapilární zónové elektroforézy pro studium enzymů