Mechanism of Selective Enzyme Inhibition through Uncompetitive Regulation of an Allosteric Agonist

Journal of the American Chemical Society
Stephen BoultonGiuseppe Melacini

Abstract

Classical uncompetitive inhibitors are potent pharmacological modulators of enzyme function. Since they selectively target enzyme-substrate complexes (E:S), their inhibitory potency is amplified by increasing substrate concentrations. Recently, an unconventional uncompetitive inhibitor, called CE3F4R, was discovered for the exchange protein activated by cAMP isoform 1 (EPAC1). Unlike conventional uncompetitive inhibitors, CE3F4R is uncompetitive with respect to an allosteric effector, cAMP, as opposed to the substrate (i.e., CE3F4R targets the E:cAMP rather than the E:S complex). However, the mechanism of CE3F4R as an uncompetitive inhibitor is currently unknown. Here, we elucidate the mechanism of CE3F4R's action using NMR spectroscopy. Due to limited solubility and line broadening, which pose major challenges for traditional structural determination approaches, we resorted to a combination of protein- and ligand-based NMR experiments to comparatively analyze EPAC mutations, inhibitor analogs, and cyclic nucleotide derivatives that trap EPAC at different stages of activation. We discovered that CE3F4R binds within the EPAC cAMP-binding domain (CBD) at a subdomain interface distinct from the cAMP binding site, acting as a wedge...Continue Reading

References

Nov 1, 1995·Journal of Biomolecular NMR·F DelaglioA Bax
Dec 18, 1998·Science·H KawasakiA M Graybiel
Jul 18, 2001·Journal of the American Chemical Society·W JahnkeM Zurini
Mar 29, 2002·Chembiochem : a European Journal of Chemical Biology·Wolfgang Jahnke
Apr 5, 2002·Journal of Medicinal Chemistry·Susan L McGovernBrian K Shoichet
Dec 7, 2002·Nature Structural Biology·Holger RehmannAlfred Wittinghofer
Sep 19, 2003·Journal of Medicinal Chemistry·Susan L McGovernBrian K Shoichet
Oct 3, 2003·Journal of Medicinal Chemistry·James SeidlerBrian K Shoichet
Mar 18, 2005·Journal of Medicinal Chemistry·Yulia Volovik FrenkelEddy Arnold
Jan 13, 2006·Nature Chemical Biology·Brian Y FengBrian K Shoichet
Sep 12, 2006·Journal of Biomolecular NMR·Mikhail ReibarkhGerhard Wagner
Nov 1, 2006·The Journal of Biological Chemistry·Rahul Das, Giuseppe Melacini
Nov 7, 2006·Trends in Biochemical Sciences·Johannes L Bos
Apr 24, 2007·Journal of Medicinal Chemistry·Brian Y FengChristopher P Austin
Oct 3, 2007·The Journal of Biological Chemistry·Kim N HaGianluigi Veglia
Dec 18, 2007·Biochemistry·Nathaniel J TraasethGianluigi Veglia
Jul 8, 2008·Acta Biochimica Et Biophysica Sinica·Xiaodong ChengFang Mei
Jul 29, 2008·Nature·Holger RehmannJohannes L Bos
Mar 19, 2009·Journal of the American Chemical Society·Alfonso De SimoneMichele Vendruscolo
May 1, 2009·The Journal of Biological Chemistry·Rahul DasGiuseppe Melacini
Mar 30, 2011·Proceedings of the National Academy of Sciences of the United States of America·Rajeevan SelvaratnamGiuseppe Melacini
Aug 30, 2011·The Journal of Biological Chemistry·Bryan VanSchouwenGiuseppe Melacini
Feb 14, 2012·Biophysical Journal·Rajeevan SelvaratnamGiuseppe Melacini
Oct 16, 2012·Molecular Pharmacology·Muayad AlmahariqXiaodong Cheng
Nov 10, 2012·The Journal of Biological Chemistry·Delphine CourilleauFrank Lezoualc'h
Nov 15, 2012·Proceedings of the National Academy of Sciences of the United States of America·Amy M Ruschak, Lewis E Kay
Nov 28, 2012·PloS One·Rajeevan SelvaratnamGiuseppe Melacini
May 9, 2013·Seminars in Cancer Biology·Chung-Jung Tsai, Ruth Nussinov
Aug 16, 2013·Proceedings of the National Academy of Sciences of the United States of America·Madoka AkimotoGiuseppe Melacini
Oct 9, 2013·Biochemical and Biophysical Research Communications·Delphine CourilleauJean-Paul Blondeau
Oct 24, 2013·Scientific Reports·Holger Rehmann
Nov 13, 2013·Proceedings of the National Academy of Sciences of the United States of America·Bin GongXiaodong Cheng
Sep 2, 2014·Progress in Biophysics and Molecular Biology·David M Dias, Alessio Ciulli

❮ Previous
Next ❯

Citations

Nov 23, 2019·Cells·Alveena AhmedGiuseppe Melacini
Jul 1, 2020·Science Advances·J A ByunG Melacini
Jun 20, 2020·Chemical Communications : Chem Comm·Madoka AkimotoGiuseppe Melacini
Apr 23, 2020·The Journal of Biological Chemistry·Jung Ah ByunGiuseppe Melacini
Aug 29, 2020·Cells·Karina FormosoJeanne Mialet-Perez
Dec 19, 2020·Computational and Structural Biotechnology Journal·Jung Ah ByunGiuseppe Melacini
Jan 18, 2021·Drug Discovery Today·Arne Christian Rufer
Jan 21, 2021·Cells·Marion LaudetteFrank Lezoualc'h
Sep 17, 2020·Science Advances·J A ByunG Melacini
Jun 12, 2021·The Journal of Physical Chemistry. B·Jinfeng HuangGiuseppe Melacini
Apr 17, 2020·Journal of Medicinal Chemistry·Hongzhao ShaoGiuseppe Melacini
May 14, 2019·The Journal of Physical Chemistry Letters·Pramodh VallurupalliShamasree Ghosh
May 11, 2019·Journal of Medicinal Chemistry·Stephen BoultonGiuseppe Melacini
Sep 4, 2020·Biophysical Journal·Stephen BoultonGiuseppe Melacini

❮ 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.