Binding and structural analyses of potent inhibitors of the human Ca2+/calmodulin dependent protein kinase kinase 2 (CAMKK2) identified from a collection of commercially-available kinase inhibitors
Abstract
Calcium/Calmodulin-dependent Protein Kinase Kinase 2 (CAMKK2) acts as a signaling hub, receiving signals from various regulatory pathways and decoding them via phosphorylation of downstream protein kinases - such as AMPK (AMP-activated protein kinase) and CAMK types I and IV. CAMKK2 relevance is highlighted by its constitutive activity being implicated in several human pathologies. However, at present, there are no selective small-molecule inhibitors available for this protein kinase. Moreover, CAMKK2 and its closest human homolog, CAMKK1, are thought to have overlapping biological roles. Here we present six new co-structures of potent ligands bound to CAMKK2 identified from a library of commercially-available kinase inhibitors. Enzyme assays confirmed that most of these compounds are equipotent inhibitors of both human CAMKKs and isothermal titration calorimetry (ITC) revealed that binding to some of these molecules to CAMKK2 is enthalpy driven. We expect our results to advance current efforts to discover small molecule kinase inhibitors selective to each human CAMKK.
- Authors:
-
[1];
[2];
[1];
[3];
[2];
[4];
- Univ. of Campinas (UNICAMP), Sao Paulo (Brazil)
- Univ. of North Carolina, Chapel Hill, NC (United States)
- Univ. of Campinas (UNICAMP), Sao Paulo (Brazil); Univ. of Oxford (United Kingdom)
- Univ. of Oxford (United Kingdom)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- USDOE Office of Science (SC); São Paulo Research Foundation (FAPESP); Brazilian National Council for Scientific and Technological Development (CNPq); National Cancer Institute (NCI); National Institutes of Health (NIH)
- OSTI Identifier:
- 1575012
- Grant/Contract Number:
- AC02-06CH11357; 2013/50724-5; 2014/50897-0; 465651/2014-3; R01CA218442; S10 RR029205
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; biochemistry; structural biology
Citation Formats
Profeta, Gerson S., dos Reis, Caio V., Santiago, André da S., Godoi, Paulo H. C., Fala, Angela M., Wells, Carrow I., Sartori, Roger, Salmazo, Anita P. T., Ramos, Priscila Z., Massirer, Katlin B., Elkins, Jonathan M., Drewry, David H., Gileadi, Opher, and Couñago, Rafael M. Binding and structural analyses of potent inhibitors of the human Ca2+/calmodulin dependent protein kinase kinase 2 (CAMKK2) identified from a collection of commercially-available kinase inhibitors. United States: N. p., 2019.
Web. doi:10.1038/s41598-019-52795-1.
Profeta, Gerson S., dos Reis, Caio V., Santiago, André da S., Godoi, Paulo H. C., Fala, Angela M., Wells, Carrow I., Sartori, Roger, Salmazo, Anita P. T., Ramos, Priscila Z., Massirer, Katlin B., Elkins, Jonathan M., Drewry, David H., Gileadi, Opher, & Couñago, Rafael M. Binding and structural analyses of potent inhibitors of the human Ca2+/calmodulin dependent protein kinase kinase 2 (CAMKK2) identified from a collection of commercially-available kinase inhibitors. United States. https://doi.org/10.1038/s41598-019-52795-1
Profeta, Gerson S., dos Reis, Caio V., Santiago, André da S., Godoi, Paulo H. C., Fala, Angela M., Wells, Carrow I., Sartori, Roger, Salmazo, Anita P. T., Ramos, Priscila Z., Massirer, Katlin B., Elkins, Jonathan M., Drewry, David H., Gileadi, Opher, and Couñago, Rafael M. Mon .
"Binding and structural analyses of potent inhibitors of the human Ca2+/calmodulin dependent protein kinase kinase 2 (CAMKK2) identified from a collection of commercially-available kinase inhibitors". United States. https://doi.org/10.1038/s41598-019-52795-1. https://www.osti.gov/servlets/purl/1575012.
@article{osti_1575012,
title = {Binding and structural analyses of potent inhibitors of the human Ca2+/calmodulin dependent protein kinase kinase 2 (CAMKK2) identified from a collection of commercially-available kinase inhibitors},
author = {Profeta, Gerson S. and dos Reis, Caio V. and Santiago, André da S. and Godoi, Paulo H. C. and Fala, Angela M. and Wells, Carrow I. and Sartori, Roger and Salmazo, Anita P. T. and Ramos, Priscila Z. and Massirer, Katlin B. and Elkins, Jonathan M. and Drewry, David H. and Gileadi, Opher and Couñago, Rafael M.},
abstractNote = {Calcium/Calmodulin-dependent Protein Kinase Kinase 2 (CAMKK2) acts as a signaling hub, receiving signals from various regulatory pathways and decoding them via phosphorylation of downstream protein kinases - such as AMPK (AMP-activated protein kinase) and CAMK types I and IV. CAMKK2 relevance is highlighted by its constitutive activity being implicated in several human pathologies. However, at present, there are no selective small-molecule inhibitors available for this protein kinase. Moreover, CAMKK2 and its closest human homolog, CAMKK1, are thought to have overlapping biological roles. Here we present six new co-structures of potent ligands bound to CAMKK2 identified from a library of commercially-available kinase inhibitors. Enzyme assays confirmed that most of these compounds are equipotent inhibitors of both human CAMKKs and isothermal titration calorimetry (ITC) revealed that binding to some of these molecules to CAMKK2 is enthalpy driven. We expect our results to advance current efforts to discover small molecule kinase inhibitors selective to each human CAMKK.},
doi = {10.1038/s41598-019-52795-1},
journal = {Scientific Reports},
number = 1,
volume = 9,
place = {United States},
year = {Mon Nov 11 00:00:00 EST 2019},
month = {Mon Nov 11 00:00:00 EST 2019}
}
Free Publicly Available Full Text
Publisher's Version of Record
Other availability
Search WorldCat to find libraries that may hold this journal
Cited by: 11 works
Citation information provided by
Web of Science
Web of Science
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
The Ca 2+ /Calmodulin/CaMKK2 Axis: Nature's Metabolic CaMshaft
journal, October 2016
- Marcelo, Kathrina L.; Means, Anthony R.; York, Brian
- Trends in Endocrinology & Metabolism, Vol. 27, Issue 10
High-throughput production of human proteins for crystallization: The SGC experience
journal, October 2010
- Savitsky, Pavel; Bray, James; Cooper, Christopher D. O.
- Journal of Structural Biology, Vol. 172, Issue 1
Overview of the CCP 4 suite and current developments
journal, March 2011
- Winn, Martyn D.; Ballard, Charles C.; Cowtan, Kevin D.
- Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4
Structural Analysis of Inhibitor Binding to CAMKK1 Identifies Features Necessary for Design of Specific Inhibitors
journal, October 2018
- Santiago, André da Silva; Couñago, Rafael M.; Ramos, Priscila Zonzini
- Scientific Reports, Vol. 8, Issue 1
Comprehensive analysis of kinase inhibitor selectivity
journal, October 2011
- Davis, Mindy I.; Hunt, Jeremy P.; Herrgard, Sanna
- Nature Biotechnology, Vol. 29, Issue 11
Integration and global analysis of isothermal titration calorimetry data for studying macromolecular interactions
journal, April 2016
- Brautigam, Chad A.; Zhao, Huaying; Vargas, Carolyn
- Nature Protocols, Vol. 11, Issue 5
Comprehensive assay of kinase catalytic activity reveals features of kinase inhibitor selectivity
journal, October 2011
- Anastassiadis, Theonie; Deacon, Sean W.; Devarajan, Karthik
- Nature Biotechnology, Vol. 29, Issue 11
Inhibition of Tumor Cell Growth, Invasion, and Metastasis by EXEL-2880 (XL880, GSK1363089), a Novel Inhibitor of HGF and VEGF Receptor Tyrosine Kinases
journal, October 2009
- Qian, F.; Engst, S.; Yamaguchi, K.
- Cancer Research, Vol. 69, Issue 20
MolProbity : all-atom structure validation for macromolecular crystallography
journal, December 2009
- Chen, Vincent B.; Arendall, W. Bryan; Headd, Jeffrey J.
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1
Calmodulin kinases: essential regulators in health and disease
journal, April 2017
- Takemoto-Kimura, Sayaka; Suzuki, Kanzo; Horigane, Shin-ichiro
- Journal of Neurochemistry, Vol. 141, Issue 6
Features and development of Coot
journal, March 2010
- Emsley, P.; Lohkamp, B.; Scott, W. G.
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4
Regulation and function of AMPK in physiology and diseases
journal, July 2016
- Jeon, Sang-Min
- Experimental & Molecular Medicine, Vol. 48, Issue 7
REFMAC 5 for the refinement of macromolecular crystal structures
journal, March 2011
- Murshudov, Garib N.; Skubák, Pavol; Lebedev, Andrey A.
- Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4
1,2,6-Thiadiazinones as Novel Narrow Spectrum Calcium/Calmodulin-Dependent Protein Kinase Kinase 2 (CaMKK2) Inhibitors
journal, May 2018
- Asquith, Christopher; Godoi, Paulo; Couñago, Rafael
- Molecules, Vol. 23, Issue 5
SFCHECK : a unified set of procedures for evaluating the quality of macromolecular structure-factor data and their agreement with the atomic model
journal, January 1999
- Vaguine, Alexei A.; Richelle, Jean; Wodak, S. J.
- Acta Crystallographica Section D Biological Crystallography, Vol. 55, Issue 1
Structure of mammalian AMPK and its regulation by ADP
journal, March 2011
- Xiao, Bing; Sanders, Matthew J.; Underwood, Elizabeth
- Nature, Vol. 472, Issue 7342
A helix scaffold for the assembly of active protein kinases
journal, September 2008
- Kornev, A. P.; Taylor, S. S.; Ten Eyck, L. F.
- Proceedings of the National Academy of Sciences, Vol. 105, Issue 38
Evidence for the Existence of Ca2+/calmodulin-Dependent Protein Kinase IV Kinase Isoforms in Rat Brain
journal, June 1996
- Okuno, S.; Kitani, T.; Fujisawa, H.
- Journal of Biochemistry, Vol. 119, Issue 6
How Much Binding Affinity Can be Gained by Filling a Cavity?
journal, February 2010
- Kawasaki, Yuko; Chufan, Eduardo E.; Lafont, Virginie
- Chemical Biology & Drug Design, Vol. 75, Issue 2
Deletion of CaMKK2 from the Liver Lowers Blood Glucose and Improves Whole-Body Glucose Tolerance in the Mouse
journal, February 2012
- Anderson, Kristin A.; Lin, Fumin; Ribar, Thomas J.
- Molecular Endocrinology, Vol. 26, Issue 2
An orally available, brain-penetrant CAMKK2 inhibitor reduces food intake in rodent model
journal, June 2018
- Price, Daniel J.; Drewry, David H.; Schaller, Lee T.
- Bioorganic & Medicinal Chemistry Letters, Vol. 28, Issue 10
Hypothalamic CaMKK2 Contributes to the Regulation of Energy Balance
journal, May 2008
- Anderson, Kristin A.; Ribar, Thomas J.; Lin, Fumin
- Cell Metabolism, Vol. 7, Issue 5
Calmodulin-Kinases: Modulators of Neuronal Development and Plasticity
journal, September 2008
- Wayman, Gary A.; Lee, Yong-Seok; Tokumitsu, Hiroshi
- Neuron, Vol. 59, Issue 6
The Ca 2+ /Calmodulin-Dependent Protein Kinase Kinase, CaMKK2, Inhibits Preadipocyte Differentiation
journal, October 2011
- Lin, Fumin; Ribar, Thomas J.; Means, Anthony R.
- Endocrinology, Vol. 152, Issue 10
STO-609, a Specific Inhibitor of the Ca 2+ /Calmodulin-dependent Protein Kinase Kinase
journal, February 2002
- Tokumitsu, Hiroshi; Inuzuka, Hiroyuki; Ishikawa, Yumi
- Journal of Biological Chemistry, Vol. 277, Issue 18
Crystal Structure of the Ca 2+ /Calmodulin-dependent Protein Kinase Kinase in Complex with the Inhibitor STO-609
journal, April 2011
- Kukimoto-Niino, Mutsuko; Yoshikawa, Seiko; Takagi, Tetsuo
- Journal of Biological Chemistry, Vol. 286, Issue 25
Forebrain-specific constitutively active CaMKKα transgenic mice show deficits in hippocampus-dependent long-term memory
journal, September 2011
- Kaitsuka, Taku; Li, Sheng-Tian; Nakamura, Kenji
- Neurobiology of Learning and Memory, Vol. 96, Issue 2
Characterization of a Ca 2+ /Calmodulin-dependent Protein Kinase Cascade : MOLECULAR CLONING AND EXPRESSION OF CALCIUM/CALMODULIN-DEPENDENT PROTEIN KINASE KINASE
journal, August 1995
- Tokumitsu, Hiroshi; Enslen, Hervé; Soderling, Thomas R.
- Journal of Biological Chemistry, Vol. 270, Issue 33
XDS
journal, January 2010
- Kabsch, Wolfgang
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2
The selectivity of protein kinase inhibitors: a further update
journal, November 2007
- Bain, Jenny; Plater, Lorna; Elliott, Matt
- Biochemical Journal, Vol. 408, Issue 3
Compensating Enthalpic and Entropic Changes Hinder Binding Affinity Optimization
journal, June 2007
- Lafont, Virginie; Armstrong, Anthony A.; Ohtaka, Hiroyasu
- Chemical Biology & Drug Design, Vol. 69, Issue 6
Molecular Cloning of Ca2+/Calmodulin-Dependent Protein Kinase Kinase
journal, July 1997
- Kitani, T.; Okuno, S.; Fujisawa, H.
- Journal of Biochemistry, Vol. 122, Issue 1
AMPK functions as an adenylate charge-regulated protein kinase
journal, March 2012
- Oakhill, Jonathan S.; Scott, John W.; Kemp, Bruce. E.
- Trends in Endocrinology & Metabolism, Vol. 23, Issue 3
Design strategies to target crystallographic waters applied to the Hsp90 molecular chaperone
journal, June 2011
- Kung, Pei-Pei; Sinnema, Piet-Jan; Richardson, Paul
- Bioorganic & Medicinal Chemistry Letters, Vol. 21, Issue 12
Phaser crystallographic software
journal, July 2007
- McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D.
- Journal of Applied Crystallography, Vol. 40, Issue 4
The Ca2+–calmodulin-dependent protein kinase cascade
journal, June 1999
- Soderling, Thomas R.
- Trends in Biochemical Sciences, Vol. 24, Issue 6