Electrophoretically mediated microanalysis with partial filling
technique and indirect or direct detection as a tool for
inhibition studies of enzymatic reaction

D 2004

Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reaction

TELNAROVÁ, Magdalena, Soňa VYTISKOVÁ, Kateřina PAPEŽOVÁ, Marta MONINCOVÁ, Zdeněk GLATZ et. al.

Basic information

Original name

Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reaction

Name in Czech

Studium inhibice enzymů kombinace metody EMMA s technikou částečného plnění kapiláry a přímou či nepřímou detekcí.

Authors

TELNAROVÁ, Magdalena (203 Czech Republic), Soňa VYTISKOVÁ (203 Czech Republic), Kateřina PAPEŽOVÁ (203 Czech Republic), Marta MONINCOVÁ (203 Czech Republic) and Zdeněk GLATZ (203 Czech Republic, guarantor)

Edition

Rakousko, Proceeding of 17th International Symposium on Microscale Separations and Capillary Electrophoresis, p. 80-80, 2004

Publisher

Tyrol Congress

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Austria

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/04:00009929

Organization unit

Faculty of Science

UT WoS

000221168700013

Keywords in English

capillary electrophoresis; microscale separations

Abstract

ORIG CZ

V originále

The study of enzymes remains of great importance to scientific community and to society in general. They are utilised in many industrial applications. The traditional roles of enzymes in food and beverage manufacturing are still in use today. In modern times, the role of enzymes in consumer products and in chemical manufacturing has expanded greatly. Perhaps one of the most exciting fields of modern enzymology is the application of enzyme inhibitors. Many of the drugs function by inhibiting specific enzymes those are associated with the disease process. Enzyme inhibitors are developed as pesticides to protect crops. Even lots of chemical warfare agents act as enzyme inhibitors. For that reason new methodological approaches are looked out for the inhibition studies of enzymatic reaction. The inhibition study of the model enzyme - haloalkane dehalogenase [EC 3.8.1.5] from Sphingomonas paucimobilis was performed by a combination of the electrophoretically mediated microanalysis with a partial filling technique and indirect or direct detection. In this set-up, the part of capillary is filled with the buffer best for the enzymatic reaction 20 mM glycine buffer (pH 8.6) whereas, the rest of the capillary is filled with the background electrolyte optimal for separation of substrates and products. Two different background electrolytes and corresponding detection approaches were used to show the versatility of the developed method. The inhibition effect of 1,2-dichloroethane on the dehalogenation of brominated substrate 1-bromobutane was studied by means of 10 mM chromate 0.1 CTAB (pH 9.2) in combination with indirect detection and 0.1 M beta-alanine HCl (pH 3.5) in combination with direct detection. For the first time the chromate background electrolyte containing tetraalkylammonium salt as an EOF modifier in combination indirect detection was used for this purpose. The method was used not only to estimate KI but also for the determination of the inhibition type. Furthermore it can be applied for the kinetic studies of the other enzymes dealing with of the high mobile inorganic or organic anions as substrates or products.

In Czech

Jedna se o studium inhibice enzymů kombinací metody EMMA s technikou částečného plnění kapiláry a přímou či nepřímou detekcí.

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

MSM 143100005, plan (intention)

Name: Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Investor: Ministry of Education, Youth and Sports of the CR, Biomolecular Structure-function Relationships and their role in the Metabolism

Citovat

TELNAROVÁ, Magdalena, Soňa VYTISKOVÁ, Kateřina PAPEŽOVÁ, Marta MONINCOVÁ and Zdeněk GLATZ. Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reaction. In Proceeding of 17th International Symposium on Microscale Separations and Capillary Electrophoresis. Rakousko: Tyrol Congress, 2004, p. 80.

@inproceedings{492939,
   author = {Telnarová, Magdalena and Vytisková, Soňa and Papežová, Kateřina and Monincová, Marta and Glatz, Zdeněk},
   address = {Rakousko},
   booktitle = {Proceeding of 17th International Symposium on Microscale Separations and Capillary Electrophoresis},
   keywords = {capillary electrophoresis; microscale separations},
   language = {eng},
   location = {Rakousko},
   pages = {80-80},
   publisher = {Tyrol Congress},
   title = {Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reaction},
   year = {2004}
}

TY  - JOUR
ID  - 492939
AU  - Telnarová, Magdalena - Vytisková, Soňa - Papežová, Kateřina - Monincová, Marta - Glatz, Zdeněk
PY  - 2004
TI  - Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reaction
PB  - Tyrol Congress
CY  - Rakousko
KW  - capillary electrophoresis
KW  - microscale separations
N2  - The study of enzymes remains of great importance to scientific community and to society in general. They are utilised in many industrial applications. The traditional roles of enzymes in food and beverage manufacturing are still in use today. In modern times, the role of enzymes in consumer products and in chemical manufacturing has expanded greatly. Perhaps one of the most exciting fields of modern enzymology is the application of enzyme inhibitors. Many of the drugs function by inhibiting specific enzymes those are associated with the disease process. Enzyme inhibitors are developed as pesticides to protect crops. Even lots of chemical warfare agents act as enzyme inhibitors. For that reason new methodological approaches are looked out for the inhibition studies of enzymatic reaction. The inhibition study of the model enzyme - haloalkane dehalogenase [EC 3.8.1.5] from Sphingomonas paucimobilis was performed by a combination of the electrophoretically mediated microanalysis with a partial filling technique and indirect or direct detection. In this set-up, the part of capillary is filled with the buffer best for the enzymatic reaction 20 mM glycine buffer (pH 8.6) whereas, the rest of the capillary is filled with the background electrolyte optimal for separation of substrates and products. Two different background electrolytes and corresponding detection approaches were used to show the versatility of the developed method. The inhibition effect of 1,2-dichloroethane on the dehalogenation of brominated substrate 1-bromobutane was studied by means of 10 mM chromate 0.1 CTAB (pH 9.2) in combination with indirect detection and 0.1 M beta-alanine HCl (pH 3.5) in combination with direct detection. For the first time the chromate background electrolyte containing tetraalkylammonium salt as an EOF modifier in combination indirect detection was used for this purpose. The method was used not only to estimate KI but also for the determination of the inhibition type. Furthermore it can be applied for the kinetic studies of the other enzymes dealing with of the high mobile inorganic or organic anions as substrates or products.
ER  -

TELNAROVÁ, Magdalena, Soňa VYTISKOVÁ, Kateřina PAPEŽOVÁ, Marta MONINCOVÁ and Zdeněk GLATZ. Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reaction. In \textit{Proceeding of 17th International Symposium on Microscale Separations and Capillary Electrophoresis}. Rakousko: Tyrol Congress, 2004, p.~80.

Detailed Information on Publication Record