Kinetic analysis of substrate competition in enzymatic reactions with β-D-galactosidase by capillary electrophoresis / dynamic frontal analysis.

Journal of Pharmaceutical and Biomedical Analysis
Masanori MineToshio Takayanagi

Abstract

Competitive inhibition between two substrates with an enzyme is investigated by capillary electrophoresis/dynamic frontal analysis (CE/DFA). Enzymatic hydrolyses of o-nitrophenyl β-D-galactopyranoside and p-nitrophenyl β-D-galactopyranoside with β-D-galactosidase were examined as a model competitive reaction. A sample solution containing the two substrates was injected into a capillary filled with a separation buffer containing an enzyme. Enzymatic hydrolysis occurred during the electrophoresis, and the products of o-nitrophenol and p-nitrophenol were continuously formed and resolved from the sample zone. Two-steps plateau signal was detected with the two-substrate solutions based on the difference in the effective electrophoretic mobility of o-nitrophenol and p-nitrophenol. Michaelis-Menten constants and inhibition constants were determined with the plateau heights. Usefulness of CE/DFA on competitive inhibition analysis is demonstrated in this study.

References

Sep 11, 1992·Journal of Chromatography·J Bao, F E Regnier
Dec 10, 1997·Journal of Chromatography. B, Biomedical Sciences and Applications·J J BaoN D Danielson
Apr 7, 2004·Journal of Chromatography. a·Sona NovákováZdenĕk Glatz
May 5, 2010·Journal of Pharmaceutical and Biomedical Analysis·Yi Fan, Gerhard K E Scriba
Jun 15, 2011·Developmental Cell·Yoosik KimStanislav Y Shvartsman
Apr 17, 2012·Analytical Chemistry·Douglas B CraigColeen Marie Q Ong-Justiniano
Sep 27, 2012·Protein Science : a Publication of the Protein Society·Douglas H JuersReuben E Huber
Jul 3, 2013·FEBS Letters·Sascha SchäubleInes Heiland
Apr 17, 2015·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mohamed Amara CamaraLi Yang
Aug 12, 2015·Molecular Pharmaceutics·Hao XuGordon L Amidon
Mar 5, 2016·Journal of Pharmaceutical and Biomedical Analysis·Annalisa D'UrzoVincenza Andrisano
Aug 25, 2018·Journal of Pharmaceutical Analysis·Mengxia Cheng, Zilin Chen
Sep 9, 2019·Biochimica Et Biophysica Acta. Proteins and Proteomics·Carlos VeraAndrés Illanes

❮ Previous
Next ❯

Citations

Sep 13, 2021·Electrophoresis·Jef RozenskiAnn Van Schepdael
Apr 13, 2021·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Junku TakaoKenji Sueyoshi

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry
Toshio TakayanagiHitoshi Mizuguchi
International Journal of Molecular Sciences
Da-Peng ZhangZhou-Sheng Yang
Biochemical and Biophysical Research Communications
J C SpainD T Gibson
Methods : a Companion to Methods in Enzymology
Srikanth GattuLisa A Holland
© 2022 Meta ULC. All rights reserved