Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

You’ve supercharged your research process with ACS and Mendeley!

STEP 1:
Click to create an ACS ID

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

MENDELEY PAIRING EXPIRED
Your Mendeley pairing has expired. Please reconnect
ACS Publications. Most Trusted. Most Cited. Most Read
My Activity
CONTENT TYPES
RETURN TO ISSUEPREVAccelerated Publicat...Accelerated PublicationNEXT

Inhibition of IMPDH by Mycophenolic Acid:  Dissection of Forward and Reverse Pathways Using Capillary Electrophoresis

View Author Information
Vertex Pharmaceuticals Incorporated, 40 Allston Street, Cambridge, Massachusetts 02139-4211
Cite this: Biochemistry 1996, 35, 22, 6990–6997
Publication Date (Web):June 4, 1996
https://doi.org/10.1021/bi9607416
Copyright © 1996 American Chemical Society

    Article Views

    657

    Altmetric

    -

    Citations

    LEARN ABOUT THESE METRICS
    Other access options

    Abstract

    The objective of this work was to contribute to the understanding of mechanisms for IMPDH inhibition. We over-expressed hamster type II IMPDH in Escherichia coli, purified the protein to apparent homogeneity, and used capillary electrophoresis to quantify enzyme turnover events accompanying inhibition by mycophenolic acid (MPA). We dissected two convergent pathways leading to MPA-inhibition; a rapid “forward” pathway beginning with substrates and linked to enzyme catalysis, and a slower “reverse” pathway apparently not involving catalysis. MPA-inhibition occurred rapidly in the forward direction by interrupting the enzyme turnover cycle, after IMP and NAD+ binding, after hydride transfer, and after NADH release. Slow inhibition, without substrate turnover, was achieved by incubating free enzyme with excess XMP and MPA. We propose that mycophenolic acid inhibits IMPDH by trapping a transient covalent product of the hydride transfer reaction (IMPDH∼XMP*) before a final hydrolysis step that precedes XMP and enzyme release in the forward reaction pathway. Understanding the ligand occupancy of the protein has also proven important for producing homogeneous, chemically defined complexes for structural studies. IMPDH samples inhibited by MPA in the forward and reverse pathways yielded similar, high-quality crystals that are currently undergoing X-ray diffraction analyses.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.

    Recommended

    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    *

     Authors to whom correspondence should be addressed. Tel:  (617) 576-3111. FAX:  (617) 576-2109. E-mail:  [email protected] and [email protected].

     Present address:  Groton School, Farmers Row, Groton, MA 01450.

     Abstract published in Advance ACS Abstracts, May 15, 1996.

    Cited By

    This article is cited by 34 publications.

    1. Magdalena Makowska-Grzyska, Youngchang Kim, Ruiying Wu, Rosemarie Wilton, Deviprasad R. Gollapalli, Ximi K. Wang, Rongguang Zhang, Robert Jedrzejczak, Jamey C. Mack, Natalia Maltseva, Rory Mulligan, T. Andrew Binkowski, Piotr Gornicki, Misty L. Kuhn, Wayne F. Anderson, Lizbeth Hedstrom, and Andrzej Joachimiak . Bacillus anthracis Inosine 5′-Monophosphate Dehydrogenase in Action: The First Bacterial Series of Structures of Phosphate Ion-, Substrate-, and Product-Bound Complexes. Biochemistry 2012, 51 (31) , 6148-6163. https://doi.org/10.1021/bi300511w
    2. Lizbeth Hedstrom. IMP Dehydrogenase: Structure, Mechanism, and Inhibition. Chemical Reviews 2009, 109 (7) , 2903-2928. https://doi.org/10.1021/cr900021w
    3. James G. Robertson. Mechanistic Basis of Enzyme-Targeted Drugs. Biochemistry 2005, 44 (15) , 5561-5571. https://doi.org/10.1021/bi050247e
    4. T. G. Murali Dhar,, Zhongqi Shen,, Junqing Guo,, Chunjian Liu,, Scott H. Watterson,, Henry H. Gu,, William J. Pitts,, Catherine A. Fleener,, Katherine A. Rouleau,, N. Z. Sherbina,, Kim W. McIntyre,, Mark R. Witmer,, Jeffrey A. Tredup,, Bang-Chi Chen,, Rulin Zhao,, Mark S. Bednarz,, Daniel L. Cheney,, John F. MacMaster,, Laura M. Miller,, Karen K. Berry,, Timothy W. Harper,, Joel C. Barrish,, Diane L. Hollenbaugh, and, Edwin J. Iwanowicz. Discovery of N-[2-[2-[[3-Methoxy-4-(5-oxazolyl)phenyl]amino]-5-oxazolyl]phenyl]-N-methyl-4- morpholineacetamide as a Novel and Potent Inhibitor of Inosine Monophosphate Dehydrogenase with Excellent in Vivo Activity. Journal of Medicinal Chemistry 2002, 45 (11) , 2127-2130. https://doi.org/10.1021/jm0105777
    5. David A. Pearlman and, Paul S. Charifson. Improved Scoring of Ligand−Protein Interactions Using OWFEG Free Energy Grids. Journal of Medicinal Chemistry 2001, 44 (4) , 502-511. https://doi.org/10.1021/jm000375v
    6. Ronald Bentley. Mycophenolic Acid:  A One Hundred Year Odyssey from Antibiotic to Immunosuppressant. Chemical Reviews 2000, 100 (10) , 3801-3826. https://doi.org/10.1021/cr990097b
    7. Paul S. Charifson,, Joseph J. Corkery,, Mark A. Murcko, and, W. Patrick Walters. Consensus Scoring:  A Method for Obtaining Improved Hit Rates from Docking Databases of Three-Dimensional Structures into Proteins. Journal of Medicinal Chemistry 1999, 42 (25) , 5100-5109. https://doi.org/10.1021/jm990352k
    8. Frank G. Whitby,, Hartmut Luecke,, Peter Kuhn,, John R. Somoza,, Jorge A. Huete-Perez,, John D. Phillips,, Christopher P. Hill,, Robert J. Fletterick, and, Ching Chung Wang. Crystal Structure of Tritrichomonas foetus Inosine-5‘-monophosphate Dehydrogenase and the Enzyme−Product Complex. Biochemistry 1997, 36 (35) , 10666-10674. https://doi.org/10.1021/bi9708850
    9. Shailesh Perdalkar, Dani Lakshman Yarlagadda, Cannanore Ganesh Pai, Shiran Shetty, Krishnamurthy Bhat. Liquid chromatographic methods in the determination of inosine monophosphate dehydrogenase enzyme activity: a review. Bioanalysis 2022, 14 (22) , 1453-1470. https://doi.org/10.4155/bio-2022-0212
    10. Amy Tarangelo, Jason Rodencal, Joon Tae Kim, Leslie Magtanong, Jonathan Z Long, Scott J Dixon. Nucleotide biosynthesis links glutathione metabolism to ferroptosis sensitivity. Life Science Alliance 2022, 5 (4) , e202101157. https://doi.org/10.26508/lsa.202101157
    11. Geoffrey A. Holdgate, Thomas D. Meek, Rachel L. Grimley. Mechanistic enzymology in drug discovery: a fresh perspective. Nature Reviews Drug Discovery 2018, 17 (2) , 115-132. https://doi.org/10.1038/nrd.2017.219
    12. Muhrez Kienana, Nadal-Desbarats Lydie, Halimi Jean-Michel, Dieme Binta, Büchler Matthias, Emond Patrick, Blasco Hélène, Le Guellec Chantal. Elucidating time-dependent changes in the urinary metabolome of renal transplant patients by a combined 1 H NMR and GC-MS approach. Molecular BioSystems 2015, 11 (9) , 2493-2510. https://doi.org/10.1039/C5MB00108K
    13. Shinuo Cao, Gabriel Oluga Aboge, Mohamad Alaa Terkawi, Mo Zhou, Yuzi Luo, Longzheng Yu, Yan Li, Younkyoung Goo, Ketsarin Kamyingkird, Tatsunori Masatani, Hiroshi Suzuki, Ikuo Igarashi, Yoshifumi Nishikawa, Xuenan Xuan. Cloning, characterization and validation of inosine 5′-monophosphate dehydrogenase of Babesia gibsoni as molecular drug target. Parasitology International 2013, 62 (2) , 87-94. https://doi.org/10.1016/j.parint.2012.10.005
    14. Enrico Rinaldelli, Alessandra Panattoni, Andrea Luvisi, Enrico Triolo. Effect of mycophenolic acid on trans-plasma membrane electron transport and electric potential in virus-infected plant tissue. Plant Physiology and Biochemistry 2012, 60 , 137-140. https://doi.org/10.1016/j.plaphy.2012.08.002
    15. Hye-Jin Shin, Soon-Ho Kwon, Ji-Myeong Park, Soon-Hyo Kwon, Kyoung-Ryul Lee, Young-Jin Kim, Sang-Hoo Lee. Quantitative determination of inosine 5'-monophosphate dehydrogenase activity in human peripheral blood mononuclear cells by ion-pair reversed-phase high-performance liquid chromatography. Analytical Science and Technology 2010, 23 (6) , 531-536. https://doi.org/10.5806/AST.2010.23.6.531
    16. Persis P. Wadia, Nancy D. Herrera, Michael M. Abecassis, Anat R. Tambur. Mycophenolic acid inhibits maturation and function of human dendritic cells and B cells. Human Immunology 2009, 70 (9) , 692-700. https://doi.org/10.1016/j.humimm.2009.05.002
    17. David A. Pearlman, B. Govinda Rao, Paul Charifson. FURSMASA: A new approach to rapid scoring functions that uses a MD‐averaged potential energy grid and a solvent‐accessible surface area term with parameters GA fit to experimental data. Proteins: Structure, Function, and Bioinformatics 2008, 71 (3) , 1519-1538. https://doi.org/10.1002/prot.21991
    18. Qingning Shu, Vasu Nair. Inosine monophosphate dehydrogenase (IMPDH) as a target in drug discovery. Medicinal Research Reviews 2008, 28 (2) , 219-232. https://doi.org/10.1002/med.20104
    19. Vasu Nair, Qingning Shu. Inosine Monophosphate Dehydrogenase as a Probe in Antiviral Drug Discovery. Antiviral Chemistry and Chemotherapy 2007, 18 (5) , 245-258. https://doi.org/10.1177/095632020701800501
    20. Melissa Swope Willis, James K. Hogan, Prakash Prabhakar, Xun Liu, Kuenhi Tsai, Yunyi Wei, Ted Fox. Investigation of protein refolding using a fractional factorial screen: A study of reagent effects and interactions. Protein Science 2005, 14 (7) , 1818-1826. https://doi.org/10.1110/ps.051433205
    21. Barry D. Kahan. Mechanisms of Pharmacologic Immune Suppression. 2003, 79-113. https://doi.org/10.1007/978-1-4615-0245-6_4
    22. Jugnu Jain, Susan J. Almquist, Angela D. Heiser, Dina Shlyakhter, Eduardo Leon, Christine Memmott, Cameron Stuver Moody, Elmar Nimmesgern, Caroline Decker. Characterization of Pharmacological Efficacy of VX-148, a New, Potent Immunosuppressive Inosine 5′-Monophosphate Dehydrogenase Inhibitor. Journal of Pharmacology and Experimental Therapeutics 2002, 302 (3) , 1272-1277. https://doi.org/10.1124/jpet.102.035659
    23. Joel A Yalowitz, Krzysztof Pankiewicz, Steven E Patterson, Hiremagalur N Jayaram. Cytotoxicity and cellular differentiation activity of methylenebis(phosphonate) analogs of tiazofurin and mycophenolic acid adenine dinucleotide in human cancer cell lines. Cancer Letters 2002, 181 (1) , 31-38. https://doi.org/10.1016/S0304-3835(02)00045-9
    24. Olga Futer, Michael D. Sintchak, Paul R. Caron, Elmar Nimmesgern, Maureen T. DeCenzo, David J. Livingston, Scott A. Raybuck. A mutational analysis of the active site of human type II inosine 5′-monophosphate dehydrogenase. Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology 2002, 1594 (1) , 27-39. https://doi.org/10.1016/S0167-4838(01)00277-1
    25. Krzysztof W Pankiewicz. Inhibitors of inosine monophosphate dehydrogenase as potential chemotherapeutic agents. Expert Opinion on Therapeutic Patents 2001, 11 (7) , 1161-1170. https://doi.org/10.1517/13543776.11.7.1161
    26. David E. Metzler, Carol M. Metzler, David J. Sauke. Metabolism of Aromatic Compounds and Nucleic Acid Bases. 2001, 1421-1471. https://doi.org/10.1016/B978-012492543-4/50028-3
    27. Michael D. Sintchak, Elmar Nimmesgern. The structure of inosine 5′-monophosphate dehydrogenase and the design of novel inhibitors. Immunopharmacology 2000, 47 (2-3) , 163-184. https://doi.org/10.1016/S0162-3109(00)00193-4
    28. Kathleen M. Kerr, Marguerite Cahoon, Daryl A. Bosco, Lizbeth Hedstrom. Monovalent Cation Activation in Escherichia coli Inosine 5′-Monophosphate Dehydrogenase. Archives of Biochemistry and Biophysics 2000, 375 (1) , 131-137. https://doi.org/10.1006/abbi.1999.1644
    29. Elmar Nimmesgern, James Black, Olga Futer, John R. Fulghum, Stephen P. Chambers, Christopher L. Brummel, Scott A. Raybuck, Michael D. Sintchak. Biochemical Analysis of the Modular Enzyme Inosine 5′-Monophosphate Dehydrogenase. Protein Expression and Purification 1999, 17 (2) , 282-289. https://doi.org/10.1006/prep.1999.1136
    30. Jonathan M. Moore. NMR techniques for characterization of ligand binding: Utility for lead generation and optimization in drug discovery. Biopolymers 1999, 51 (3) , 221-243. https://doi.org/10.1002/(SICI)1097-0282(1999)51:3<221::AID-BIP5>3.0.CO;2-9
    31. Theodore M. Sievers, Stephen J. Rossi, Rafik M. Ghobrial, Edgar Arriola, Pamela Nishimura, Marv Kawano, Curtis D. Holt. Mycophenolate Mofetil. Pharmacotherapy: The Journal of Human Pharmacology and Drug Therapy 1997, 17 (6) , 1178-1197. https://doi.org/10.1002/j.1875-9114.1997.tb03082.x
    32. Thalia Farazi, Joshua Leichman, Thanawath Harris, Marguerite Cahoon, Lizbeth Hedstrom. Isolation and Characterization of Mycophenolic Acid-resistant Mutants of Inosine-5′-monophosphate Dehydrogenase. Journal of Biological Chemistry 1997, 272 (2) , 961-965. https://doi.org/10.1074/jbc.272.2.961
    33. Elmar Nimmesgern, Ted Fox, Mark A. Fleming, John A. Thomson. Conformational Changes and Stabilization of Inosine 5′-Monophosphate Dehydrogenase Associated with Ligand Binding and Inhibition by Mycophenolic Acid. Journal of Biological Chemistry 1996, 271 (32) , 19421-19427. https://doi.org/10.1074/jbc.271.32.19421
    34. Michael D. Sintchak, Mark A. Fleming, Olga Futer, Scott A. Raybuck, Stephen P. Chambers, Paul R. Caron, Mark A. Murcko, Keith P. Wilson. Structure and Mechanism of Inosine Monophosphate Dehydrogenase in Complex with the Immunosuppressant Mycophenolic Acid. Cell 1996, 85 (6) , 921-930. https://doi.org/10.1016/S0092-8674(00)81275-1