ACS Publications. Most Trusted. Most Cited. Most Read
My Activity
CONTENT TYPES

Structural Basis of RasGRP Binding to High-Affinity PKC Ligands

View Author Information
Drug Discovery Program, Department of Neurology, Georgetown University Medical Center, 3970 Reservoir Road, Washington, D.C. 20007, Molecular Mechanisms of Tumor Promotion Section, National Cancer Institute, Bethesda, Maryland 20892, 123, State Key Laboratory of Bio-organic and Natural Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 354 Fenglin Lu, Shanghai 200032, China, and Department of Biochemistry, University of Alberta, Edmonton, Alberta T6G 2H7, Canada
Cite this: J. Med. Chem. 2002, 45, 4, 853–860
Publication Date (Web):January 22, 2002
https://doi.org/10.1021/jm010422z
Copyright © 2002 American Chemical Society

    Article Views

    550

    Altmetric

    -

    Citations

    LEARN ABOUT THESE METRICS
    Other access options

    Abstract

    Abstract Image

    The Ras guanyl releasing protein RasGRP belongs to the CDC25 class of guanyl nucleotide exchange factors that regulate Ras-related GTPases. These GTPases serve as switches for the propagation and divergence of signaling pathways. One interesting feature of RasGRP is the presence of a C-terminal C1 domain, which has high homology to the PKC C1 domain and binds to diacylglycerol (DAG) and phorbol esters. RasGRP thus represents a novel, non-kinase phorbol ester receptor. In this paper, we investigate the binding of indolactam(V) (ILV), 7-(n-octyl)-ILV, 8-(1-decynyl)benzolactam(V) (benzolactam), and 7-methoxy-8-(1-decynyl)benzolactam(V) (methoxylated benzolactam) to RasGRP through both experimental binding assays and molecular modeling studies. The binding affinities of these lactams to RasGRP are within the nanomolar range. Homology modeling was used to model the structure of the RasGRP C1 domain (C1-RasGRP), which was subsequently used to model the structures of C1-RasGRP in complex with these ligands and phorbol 13-acetate using a computational docking method. The structural model of C1-RasGRP exhibits a folding pattern that is nearly identical to that of C1b-PKCδ and is comprised of three antiparallel-strand β-sheets capped against a C-terminal α-helix. Two loops A and B comprising residues 8−12 and 21−27 form a binding pocket that has some positive charge character. The ligands phorbol 13-acetate, benzolactam, and ILV are recognized by C1-RasGRP through a number of hydrogen bonds with loops A and B. In the models of C1-RasGRP in complex with phorbol 13-acetate, benzolactam, and ILV, common hydrogen bonds are formed with two residues Thr12 and Leu21, whereas other hydrogen bond interactions are unique for each ligand. Furthermore, our modeling results suggest that the shallower insertion of ligands into the binding pocket of C1-RasGRP compared to C1b-PKCδ may be due to the presence of Phe rather than Leu at position 20 in C1-RasGRP. Taken together, our experimental and modeling studies provide us with a better understanding of the structural basis of the binding of PKC ligands to the novel phorbol ester receptor RasGRP.

    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.

     Georgetown University Medical Center.

     Current address:  NeuroLogic Incorporation, 9700 Great Seneca Highway, Rockville, MD 20850.

    §

     Shanghai Institute of Organic Chemistry.

     National Cancer Institute.

     University of Alberta.

    *

     To whom correspondence should be addressed. For A.P.K.:  phone, 202-687-0686; fax, 202-687-5065; e-mail, [email protected]. For S.W.:  phone, 202-687-2028; fax, 202-687-4032; e-mail, wangs@ giccs.georgetown.edu.

    Cited By

    This article is cited by 19 publications.

    1. Joydip Das and Ghazi M. Rahman . C1 Domains: Structure and Ligand-Binding Properties. Chemical Reviews 2014, 114 (24) , 12108-12131. https://doi.org/10.1021/cr300481j
    2. Uri Mbonye, Jonathan Karn. The cell biology of HIV-1 latency and rebound. Retrovirology 2024, 21 (1) https://doi.org/10.1186/s12977-024-00639-w
    3. Noah Isakov. Protein kinase C (PKC) isoforms in cancer, tumor promotion and tumor suppression. Seminars in Cancer Biology 2018, 48 , 36-52. https://doi.org/10.1016/j.semcancer.2017.04.012
    4. Damon S. Perez, Robert J. Handa, Raymond S. H. Yang, Julie A. Campain. Gene expression changes associated with altered growth and differentiation in benzo[a]pyrene or arsenic exposed normal human epidermal keratinocytes. Journal of Applied Toxicology 2008, 28 (4) , 491-508. https://doi.org/10.1002/jat.1301
    5. Danielle J. Vugts, Halil Aktas, Kanar Al‐Mafraji, Frans J. J. de Kanter, Eelco Ruijter, Marinus B. Groen, Romano V. A. Orru. Synthesis of 3′‐Deoxyribolactones using a Hydrolysis‐Induced Lactonization Cascade Reaction of Epoxy Cyanohydrins. European Journal of Organic Chemistry 2008, 2008 (8) , 1336-1339. https://doi.org/10.1002/ejoc.200701152
    6. Joanne E. Johnson, Rebecca E. Goulding, Ziwei Ding, Amir Partovi, Kira V. Anthony, Nadine Beaulieu, Ghazaleh Tazmini, Rosemary B. Cornell, Robert J. Kay. Differential membrane binding and diacylglycerol recognition by C1 domains of RasGRPs. Biochemical Journal 2007, 406 (2) , 223-236. https://doi.org/10.1042/BJ20070294
    7. Nadine Beaulieu, Bari Zahedi, Rebecca E. Goulding, Ghazaleh Tazmini, Kira V. Anthony, Stephanie L. Omeis, Danielle R. de Jong, Robert J. Kay, . Regulation of RasGRP1 by B Cell Antigen Receptor Requires Cooperativity between Three Domains Controlling Translocation to the Plasma Membrane. Molecular Biology of the Cell 2007, 18 (8) , 3156-3168. https://doi.org/10.1091/mbc.e06-10-0932
    8. Juan C. Gómez-Fernández, Senena Corbalán-García. Diacylglycerols, multivalent membrane modulators. Chemistry and Physics of Lipids 2007, 148 (1) , 1-25. https://doi.org/10.1016/j.chemphyslip.2007.04.003
    9. Kaoru Goto, Yasukazu Hozumi, Tomoyuki Nakano, Sachiko S. Saino, Hisatake Kondo. Cell Biology and Pathophysiology of the Diacylglycerol Kinase Family: Morphological Aspects in Tissues and Organs. 2007, 25-63. https://doi.org/10.1016/S0074-7696(07)64002-9
    10. Shujie Han, Stewart M. Knoepp, Mark A. Hallman, Kathryn E. Meier. RasGRP1 Confers the Phorbol Ester-Sensitive Phenotype to EL4 Lymphoma Cells. Molecular Pharmacology 2007, 71 (1) , 314-322. https://doi.org/10.1124/mol.106.028639
    11. Jason J. Coughlin, Stacey L. Stang, Nancy A. Dower, James C. Stone. RasGRP1 and RasGRP3 Regulate B Cell Proliferation by Facilitating B Cell Receptor-Ras Signaling. The Journal of Immunology 2005, 175 (11) , 7179-7184. https://doi.org/10.4049/jimmunol.175.11.7179
    12. Bin Chen, Pavel A. Petukhov, Mira Jung, Alfredo Velena, Elena Eliseeva, Anatoly Dritschilo, Alan P. Kozikowski. Chemistry and biology of mercaptoacetamides as novel histone deacetylase inhibitors. Bioorganic & Medicinal Chemistry Letters 2005, 15 (5) , 1389-1392. https://doi.org/10.1016/j.bmcl.2005.01.006
    13. Gregory M. Springett, Hiroaki Kawasaki, David R. Spriggs. Non‐kinase second‐messenger signaling: new pathways with new promise. BioEssays 2004, 26 (7) , 730-738. https://doi.org/10.1002/bies.20057
    14. Chaya Brodie, Rivka Steinhart, Gila Kazimirsky, Hadara Rubinfeld, Tehila Hyman, Jolene N. Ayres, Gang Min Hur, Attila Toth, Dazhi Yang, Susan H. Garfield, James C. Stone, Peter M. Blumberg. PKCδ Associates with and Is Involved in the Phosphorylation of RasGRP3 in Response to Phorbol Esters. Molecular Pharmacology 2004, 66 (1) , 76-84. https://doi.org/10.1124/mol.66.1.76
    15. Silvia Carrasco, Isabel Merida. Diacylglycerol-dependent Binding Recruits PKCθ and RasGRP1 C1 Domains to Specific Subcellular Localizations in Living T Lymphocytes. Molecular Biology of the Cell 2004, 15 (6) , 2932-2942. https://doi.org/10.1091/mbc.e03-11-0844
    16. Mariía J. Caloca, José L. Zugaza, Miguel Vicente-Manzanares, Francisco Sánchez-Madrid, Xosé R. Bustelo. F-actin-dependent Translocation of the Rap1 GDP/GTP Exchange Factor RasGRP2. Journal of Biological Chemistry 2004, 279 (19) , 20435-20446. https://doi.org/10.1074/jbc.M313013200
    17. Sihem Madani, Aziz Hichami, Mustapha Charkaoui-Malki, Naim A. Khan. Diacylglycerols Containing Omega 3 and Omega 6 Fatty Acids Bind to RasGRP and Modulate MAP Kinase Activation. Journal of Biological Chemistry 2004, 279 (2) , 1176-1183. https://doi.org/10.1074/jbc.M306252200
    18. ChengFeng Yang, Marcelo G Kazanietz. Divergence and complexities in DAG signaling: looking beyond PKC. Trends in Pharmacological Sciences 2003, 24 (11) , 602-608. https://doi.org/10.1016/j.tips.2003.09.003
    19. Christine Teixeira, Stacey L. Stang, Yong Zheng, Naomi S. Beswick, James C. Stone. Integration of DAG signaling systems mediated by PKC-dependent phosphorylation of RasGRP3. Blood 2003, 102 (4) , 1414-1420. https://doi.org/10.1182/blood-2002-11-3621

    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