Colorimetric coupled enzyme assay for gamma-glutamyltransferase activity using glutathione as substrate

Journal of Biochemical and Biophysical Methods
Antonella Del CorsoUmberto Mura

Abstract

A colorimetric coupled enzyme assay for the determination of gamma-glutamyltransferase (GGT) activity using glutathione as substrate is described. The cysteine released from glutathione upon sequential action of GGT and leucine aminopeptidase is spectrophotometrically detected through its reaction with ninhydrin at 100 degrees C in acidic conditions. The method was applied to the determination of the activity of both bovine kidney and human serum GGT. In the described assay conditions with final GGT concentrations ranging from 0.18 to 4 mU/ml, a linear relationship between produced cysteine and incubation times up to 90 min was observed. When a standard chromogenic assay for GGT using L-gamma-glutamyl-3-carboxy-4-nitroanilide as substrate and the proposed assay were applied on the same serum sample a linear relationship between the two method was observed. Since the use of GSH as substrate, the proposed method can be usefully adopted for enzymological studies on GGT-related enzymes, a class of enzymes which is still waiting to be characterized.

References

Dec 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·O W GriffithA Meister
Jul 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·N HeisterkampJ Groffen
Sep 25, 1981·Molecular and Cellular Biochemistry·S S Tate, A Meister
Oct 1, 1984·Journal of Bacteriology·R NakayamaT Tochikura
Jan 5, 1999·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·B del BelloE Maellaro
Nov 18, 2000·The Journal of Biological Chemistry·D R KarpP E Lipsky
Sep 21, 2001·Critical Reviews in Clinical Laboratory Sciences·J B Whitfield
Feb 14, 2002·Archives of Biochemistry and Biophysics·Antonella Del CorsoUmberto Mura
Oct 30, 2003·The Biochemical Journal·Mario CappielloUmberto Mura
Sep 7, 2004·Free Radical Research·Duk-Hee LeeDavid R Jacobs

❮ Previous
Next ❯

Citations

Dec 4, 2012·Analytical and Bioanalytical Chemistry·Mario CappielloUmberto Mura
Nov 13, 2008·Archives of Biochemistry and Biophysics·Valeria AngeliEmilia Bramanti
Mar 17, 2009·Biochemical and Biophysical Research Communications·Bjorn VergauwenBart Devreese
Jun 18, 2009·Archives of Biochemistry and Biophysics·Daniela GiustariniRanieri Rossi
Jun 14, 2017·Analytical Chemistry·Zijuan HaiGaolin Liang
Sep 28, 2018·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Shaman LuoPhilip Lazarus

❮ 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.