Multifunctional role of Tyr 108 in the catalytic mechanism of human glutathione transferase P1-1. Crystallographic and kinetic studies on the Y108F mutant enzyme

Biochemistry
M Lo BelloG Ricci

Abstract

The possible role of the hydroxyl group of Tyr 108 in the catalytic mechanism of human glutathione transferase P1-1 has been investigated by means of site-directed mutagenesis, steady-state kinetic analysis, and crystallographic studies. Three representative cosubstrates have been used, i.e. ethacrynic acid, 7-chloro-4-nitrobenz-2-oxa-1,3-diazole, and 1-chloro-2,4-dinitrobenzene. In the presence of ethacrynic acid, the enzyme follows a rapid equilibrium random bi-bi mechanism with a rate-limiting step which occurs after the addition of the substrates and before the release of products. The replacement of Tyr 108 with Phe yields a 14-fold decrease of k(cat), while it does not change appreciably the affinity of the H site for the substrate. In this case, it would appear that the role of the hydroxyl function is to stabilize the transition state for the chemical step, i.e. the Michael addition of GSH to the electrophilic substrate. Crystallographic data are compatible with this conclusion showing the hydroxyl group of Y108 in hydrogen bonding distance of the ketone moiety of ethacrynic acid [Oakley, A. J., Rossjohn, J., Lo Bello, M., Caccuri, A. M., Federici, G., & Parker, M. W. (1997) Biochemistry 36, 576-585]. Moreover, no struc...Continue Reading

References

Feb 15, 1992·The Biochemical Journal·B MannervikW R Pearson
May 20, 1990·Journal of Molecular Biology·M W ParkerG Federici
Sep 14, 1989·Biochimica Et Biophysica Acta·K M Ivanetich, R D Goold
Jul 10, 1993·Biochimica Et Biophysica Acta·S AwasthiG A Ansari
May 1, 1959·Archives of Biochemistry and Biophysics·G L ELLMAN

❮ Previous
Next ❯

Citations

Sep 6, 2003·International Journal of Biological Macromolecules·Montserrat Andújar-SánchezVicente Jara-Pérez
Apr 21, 1999·Biochimica Et Biophysica Acta·T BucciarelliC Di Ilio
Jul 20, 2002·The International Journal of Biochemistry & Cell Biology·Tonino BucciarelliCarmine Di Ilio
Nov 18, 1998·Current Opinion in Chemical Biology·R N Armstrong
Mar 24, 1999·Archives of Biochemistry and Biophysics·N E PettigrewR F Colman
Mar 25, 2011·Drug Metabolism Reviews·Aaron Oakley
Sep 2, 2008·Archives of Biochemistry and Biophysics·Tassanee Lerksuthirat, Albert J Ketterman
Aug 9, 2008·International Journal of Cancer. Journal International Du Cancer·Joachim MüllerAndrew Hemphill
Jun 12, 2010·Journal of Molecular Recognition : JMR·Indalecio Quesada-SorianoLuis García-Fuentes
Sep 26, 2009·Protein Science : a Publication of the Protein Society·Indalecio Quesada-SorianoLuis García-Fuentes
Jun 11, 2005·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·Périne DoyenFrançois Rodius
Mar 4, 2010·The Biochemical Journal·Jantana WongsantichonAlbert J Ketterman
Oct 6, 2012·Biochimica Et Biophysica Acta·Marcel Deponte
Nov 14, 1998·Biochemical and Biophysical Research Communications·M NuccetelliM Lo Bello
Apr 27, 2016·Molecular BioSystems·L A Crawford, E Weerapana
Jan 11, 2005·The Journal of Biological Chemistry·Markus PerbandtEva Liebau
Aug 14, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Dorota KrasowskaLuca Sancineto
Nov 8, 2008·Cancer Epidemiology, Biomarkers & Prevention : a Publication of the American Association for Cancer Research, Cosponsored by the American Society of Preventive Oncology·Alessandra PrimaveraMario Lo Bello
Nov 11, 2019·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Ivan MihaljevićTvrtko Smital
Dec 5, 2003·Journal of the American Chemical Society·Diana S HamiltonDonald J Creighton

❮ 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.