Application of Capillary Zone Electrophoresis for the Study of
Enzymes

D 2003

Application of Capillary Zone Electrophoresis for the Study of Enzymes

GLATZ, Zdeněk, Soňa NOVÁKOVÁ and Magdalena TELNAROVÁ

Basic information

Original name

Application of Capillary Zone Electrophoresis for the Study of Enzymes

Authors

GLATZ, Zdeněk (203 Czech Republic, guarantor), Soňa NOVÁKOVÁ (203 Czech Republic) and Magdalena TELNAROVÁ (203 Czech Republic)

Edition

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

Publisher

MCI Paris France

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

France

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/03:00008124

Organization unit

Faculty of Science

Keywords in English

CZE; Enzymes

Abstract

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.

Links

GA203/03/1125, research and development project

Name: Využití kapilární zónové elektroforézy pro studium enzymů

Investor: Czech Science Foundation, Application of capillary zone electrophoresis for the study of enzymes

Citovat

GLATZ, Zdeněk, Soňa NOVÁKOVÁ and Magdalena TELNAROVÁ. Application of Capillary Zone Electrophoresis for the Study of Enzymes. In Proceeding of 27th Symposium on HPLC and Related Techniques. Nice, France: MCI Paris France, 2003, p. 133-133.

@inproceedings{488015,
   author = {Glatz, Zdeněk and Nováková, Soňa and Telnarová, Magdalena},
   address = {Nice, France},
   booktitle = {Proceeding of 27th Symposium on HPLC and Related Techniques},
   keywords = {CZE; Enzymes},
   language = {eng},
   location = {Nice, France},
   pages = {133-133},
   publisher = {MCI Paris France},
   title = {Application of Capillary Zone Electrophoresis for the Study of Enzymes},
   year = {2003}
}

TY  - JOUR
ID  - 488015
AU  - Glatz, Zdeněk - Nováková, Soňa - Telnarová, Magdalena
PY  - 2003
TI  - Application of Capillary Zone Electrophoresis for the Study of Enzymes
PB  - MCI Paris France
CY  - Nice, France
KW  - CZE
KW  - Enzymes
N2  - 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.
ER  -

GLATZ, Zdeněk, Soňa NOVÁKOVÁ and Magdalena TELNAROVÁ. Application of Capillary Zone Electrophoresis for the Study of Enzymes. In \textit{Proceeding of 27th Symposium on HPLC and Related Techniques}. Nice, France: MCI Paris France, 2003, p.~133-133.

Detailed Information on Publication Record