Detailed Information on Publication Record
2004
Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reactions
TELNAROVÁ, Magdaléna, Soňa VYTISKOVÁ, Marta MONINCOVÁ and Zdeněk GLATZBasic information
Original name
Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reactions
Name in Czech
Studium inhibice enzymů kombinací metody EMMA s technikou částečného plnění kapiláry a přímou či nepřímou detekcí.
Authors
TELNAROVÁ, Magdaléna (203 Czech Republic), Soňa VYTISKOVÁ (203 Czech Republic), Marta MONINCOVÁ (203 Czech Republic) and Zdeněk GLATZ (203 Czech Republic, guarantor)
Edition
Electrophoresis, Weinheim, VHC, 2004, 0173-0835
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10600 1.6 Biological sciences
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
Impact factor
Impact factor: 3.743
RIV identification code
RIV/00216224:14310/04:00009987
Organization unit
Faculty of Science
UT WoS
000221168700013
Keywords in English
inhibition; EMMA; partial filling technique; direct or indirect detection
Změněno: 19/5/2009 18:44, prof. RNDr. Zdeněk Glatz, CSc.
V originále
The inhibition study of the model enzyme haloalkane dehalogenase 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 suitable 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 mM cetyltrimethylammonium bromide (pH 9.2) in combination with an indirect detection or 0.1 M beta-alanine - hydrochloric acid (pH 3.5) in combination with a direct detection. The method was used not only to estimate inhibition constant Ki 0.44 mM by means of indirect detection method and 0.63 mM by means of direct detection method but also to determine the inhibition type. Compared to spectrophotometric and other discontinuous assays, the method is rapid, can be automated, and requires only small amount of reagents that is especially important in the case of enzymes and inhibitors. Consequently it has a great potential for such determinations in other enzyme-inhibitor systems.
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 |
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| MSM 143100005, plan (intention) |
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