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

J 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 a Zdeněk GLATZ

Základní údaje

Originální název

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

Název česky

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

Autoři

TELNAROVÁ, Magdaléna (203 Česká republika), Soňa VYTISKOVÁ (203 Česká republika), Marta MONINCOVÁ (203 Česká republika) a Zdeněk GLATZ (203 Česká republika, garant)

Vydání

Electrophoresis, Weinheim, VHC, 2004, 0173-0835

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 3.743

Kód RIV

RIV/00216224:14310/04:00009987

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000221168700013

Klíčová slova anglicky

inhibition; EMMA; partial filling technique; direct or indirect detection

Štítky

direct or indirect detection, EMMA, inhibition, partial filling technique

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

Anotace

ORIG CZ

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.

Česky

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

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

MSM 143100005, záměr

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

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Citovat

TELNAROVÁ, Magdaléna, Soňa VYTISKOVÁ, Marta MONINCOVÁ a Zdeněk GLATZ. Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reactions. Electrophoresis. Weinheim: VHC, 2004, roč. 24, 7-8, s. 1028-1033. ISSN 0173-0835.

@article{554086,
   author = {Telnarová, Magdaléna and Vytisková, Soňa and Monincová, Marta and Glatz, Zdeněk},
   article_location = {Weinheim},
   article_number = {7-8},
   keywords = {inhibition; EMMA; partial filling technique; direct or indirect detection},
   language = {eng},
   issn = {0173-0835},
   journal = {Electrophoresis},
   title = {Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reactions},
   volume = {24},
   year = {2004}
}

TY  - JOUR
ID  - 554086
AU  - Telnarová, Magdaléna - Vytisková, Soňa - 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 reactions
JF  - Electrophoresis
VL  - 24
IS  - 7-8
SP  - 1028
EP  - 1028
PB  - VHC
SN  - 01730835
KW  - inhibition
KW  - EMMA
KW  - partial filling technique
KW  - direct or indirect detection
N2  - 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.
ER  -

TELNAROVÁ, Magdaléna, Soňa VYTISKOVÁ, Marta MONINCOVÁ a Zdeněk GLATZ. Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reactions. \textit{Electrophoresis}. Weinheim: VHC, 2004, roč.~24, 7-8, s.~1028-1033. ISSN~0173-0835.

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