Electrophoresis. 2003 Mar;24(6):1111-8.

Electrophoretically mediated reaction of glycosidases at a nanoliter scale.

Source

Mitsubishi Kagaku Institute of Life Sciences (MITILS), Tokyo, Japan.

Abstract

We have investigated electrophoretically mediated microanalysis (EMMA) for the assay of a native glyco-enzyme. As a representative of this class of enzyme, beta-glucosidase was selected, and the reaction was analyzed. Our EMMA was based on the plug-plug interaction of enzyme and substrate plugs, which is essential to reduce quantities of materials. Furthermore, we have addressed the problem of incompatibility of the enzymatic reaction and separation of the reactants. As a result, EMMA of native glycosidase was achieved with a reaction volume of approximately 20 nL and the Michaelis constant was estimated according to the Lineweaver-Burk plot. The current method may have advantages over traditional assay methods, especially in terms of the amount of enzyme (ng order) and substrate (pmol order) required for a reaction.

PMID:: 12658703; [PubMed - indexed for MEDLINE]

MeSH Terms, Substances

MeSH Terms

Substances

beta-Glucosidase

LinkOut - more resources

Full Text Sources

Supplemental Content

Save items

Related citations in PubMed

Electrophoretically mediated microscale reaction of glycosidases: kinetic analysis of some glycosidases at the nanoliter scale. [Carbohydr Res. 2002]
Electrophoretically mediated microscale reaction of glycosidases: kinetic analysis of some glycosidases at the nanoliter scale.
Kanie Y, Kanie O. Carbohydr Res. 2002 Oct 11; 337(19):1757-62.
Determination of the kinetic parameters of adenosine deaminase by electrophoretically mediated microanalysis. [Electrophoresis. 1996]
Determination of the kinetic parameters of adenosine deaminase by electrophoretically mediated microanalysis.
Saevels J, Van Schepdael A, Hoogmartens J. Electrophoresis. 1996 Jul; 17(7):1222-7.
Michaelis-Menten analysis of bovine plasma amine oxidase by capillary electrophoresis using electrophoretically mediated microanalysis in a partially filled capillary. [Electrophoresis. 2001]
Michaelis-Menten analysis of bovine plasma amine oxidase by capillary electrophoresis using electrophoretically mediated microanalysis in a partially filled capillary.
Van Dyck S, Van Schepdael A, Hoogmartens J. Electrophoresis. 2001 Apr; 22(7):1436-42.
Review Electrophoretically mediated microanalysis. [J Chromatogr A. 2004]
Review Electrophoretically mediated microanalysis.
Nováková S, Van Dyck S, Van Schepdael A, Hoogmartens J, Glatz Z. J Chromatogr A. 2004 Apr 2; 1032(1-2):173-84.
Review Advances in capillary electrophoretically mediated microanalysis. [Electrophoresis. 2003]
Review Advances in capillary electrophoretically mediated microanalysis.
Van Dyck S, Kaale E, Nováková S, Glatz Z, Hoogmartens J, Van Schepdael A. Electrophoresis. 2003 Dec; 24(22-23):3868-78.

See reviews... See all...

Cited by 1 PubMed Central article

A capillary electrophoresis method for the characterization of ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) and the analysis of inhibitors by in-capillary enzymatic microreaction. [Purinergic Signal. 2005]
A capillary electrophoresis method for the characterization of ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) and the analysis of inhibitors by in-capillary enzymatic microreaction.
Iqbal J, Vollmayer P, Braun N, Zimmermann H, Müller CE. Purinergic Signal. 2005 Dec; 1(4):349-58. Epub 2005 Dec 3.

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
Substance (MeSH Keyword)
PubChem chemical substance (submitted) records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Cited in PMC
Full-text articles in the PubMed Central Database that cite the current articles.

Recent activity

Clear Turn Off Turn On

Electrophoretically mediated reaction of glycosidases at a nanoliter scale.
Electrophoretically mediated reaction of glycosidases at a nanoliter scale.
Electrophoresis. 2003 Mar ;24(6):1111-8.

PubMed
Anterior gastropexy may reduce the recurrence rate after laparoscopic paraesopha...
Anterior gastropexy may reduce the recurrence rate after laparoscopic paraesophageal hernia repair.
Surg Endosc. 2003 Jul ;17(7):1036-41. Epub 2003 Mar 28 .

PubMed
Alzheimer's disease: the cholesterol connection.
Alzheimer's disease: the cholesterol connection.
Nat Neurosci. 2003 Apr ;6(4):345-51.

PubMed
[Intestinal ganglioneuromatosis diagnosed in adult patients].
[Intestinal ganglioneuromatosis diagnosed in adult patients].
Gastroenterol Clin Biol. 2003 Feb ;27(2):219-24.

PubMed
Primary prostatic endodermal sinus tumor (yolk sac tumor) combined with a small ...
Primary prostatic endodermal sinus tumor (yolk sac tumor) combined with a small focal seminoma.
Am J Surg Pathol. 2003 Apr ;27(4):554-9.

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Result Filters

Display Settings:

Send to: