3feg Citations

Crystal structures of human choline kinase isoforms in complex with hemicholinium-3: single amino acid near the active site influences inhibitor sensitivity.

OpenAccess logo J. Biol. Chem. 285 16330-40 (2010)
Related entries: 3g15, 3lq3

Cited: 23 times
EuropePMC logo PMID: 20299452

Abstract

Human choline kinase (ChoK) catalyzes the first reaction in phosphatidylcholine biosynthesis and exists as ChoKalpha (alpha1 and alpha2) and ChoKbeta isoforms. Recent studies suggest that ChoK is implicated in tumorigenesis and emerging as an attractive target for anticancer chemotherapy. To extend our understanding of the molecular mechanism of ChoK inhibition, we have determined the high resolution x-ray structures of the ChoKalpha1 and ChoKbeta isoforms in complex with hemicholinium-3 (HC-3), a known inhibitor of ChoK. In both structures, HC-3 bound at the conserved hydrophobic groove on the C-terminal lobe. One of the HC-3 oxazinium rings complexed with ChoKalpha1 occupied the choline-binding pocket, providing a structural explanation for its inhibitory action. Interestingly, the HC-3 molecule co-crystallized with ChoKbeta was phosphorylated in the choline binding site. This phosphorylation, albeit occurring at a very slow rate, was confirmed experimentally by mass spectroscopy and radioactive assays. Detailed kinetic studies revealed that HC-3 is a much more potent inhibitor for ChoKalpha isoforms (alpha1 and alpha2) compared with ChoKbeta. Mutational studies based on the structures of both inhibitor-bound ChoK complexes demonstrated that Leu-401 of ChoKalpha2 (equivalent to Leu-419 of ChoKalpha1), or the corresponding residue Phe-352 of ChoKbeta, which is one of the hydrophobic residues neighboring the active site, influences the plasticity of the HC-3-binding groove, thereby playing a key role in HC-3 sensitivity and phosphorylation.

Reviews citing this publication (8)

  1. Involvement of human choline kinase alpha and beta in carcinogenesis: a different role in lipid metabolism and biological functions. Gallego-Ortega D, Gómez del Pulgar T, Valdés-Mora F, Cebrián A, Lacal JC. Adv. Enzyme Regul. 51 183-194 (2011)
  2. Choline kinase alpha-Putting the ChoK-hold on tumor metabolism. Arlauckas SP, Popov AV, Delikatny EJ. Prog. Lipid Res. 63 28-40 (2016)
  3. Approved and Experimental Small-Molecule Oncology Kinase Inhibitor Drugs: A Mid-2016 Overview. Fischer PM. Med Res Rev 37 314-367 (2017)
  4. Molecular structure and differential function of choline kinases CHKα and CHKβ in musculoskeletal system and cancer. Chen X, Qiu H, Wang C, Yuan Y, Tickner J, Xu J, Zou J. Cytokine Growth Factor Rev. 33 65-72 (2017)
  5. Molecular structure and differential function of choline kinases CHKα and CHKβ in musculoskeletal system and cancer. Chen X, Qiu H, Wang C, Yuan Y, Tickner J, Xu J, Zou J. Cytokine Growth Factor Rev. 33 65-72 (2017)
  6. Approved and Experimental Small-Molecule Oncology Kinase Inhibitor Drugs: A Mid-2016 Overview. Fischer PM. Med Res Rev 37 314-367 (2017)
  7. Choline kinase alpha-Putting the ChoK-hold on tumor metabolism. Arlauckas SP, Popov AV, Delikatny EJ. Prog. Lipid Res. 63 28-40 (2016)
  8. Involvement of human choline kinase alpha and beta in carcinogenesis: a different role in lipid metabolism and biological functions. Gallego-Ortega D, Gómez del Pulgar T, Valdés-Mora F, Cebrián A, Lacal JC. Adv. Enzyme Regul. 51 183-194 (2011)

Articles citing this publication (15)

  1. Combined 5-FU and ChoKα inhibitors as a new alternative therapy of colorectal cancer: evidence in human tumor-derived cell lines and mouse xenografts. de la Cueva A, Ramírez de Molina A, Alvarez-Ayerza N, Ramos MA, Cebrián A, Del Pulgar TG, Lacal JC. PLoS ONE 8 e64961 (2013)
  2. The mechanism of allosteric coupling in choline kinase α1 revealed by the action of a rationally designed inhibitor. Sahún-Roncero M, Rubio-Ruiz B, Saladino G, Conejo-García A, Espinosa A, Velázquez-Campoy A, Gervasio FL, Entrena A, Hurtado-Guerrero R. Angew. Chem. Int. Ed. Engl. 52 4582-4586 (2013)
  3. Novel 4-amino bis-pyridinium and bis-quinolinium derivatives as choline kinase inhibitors with antiproliferative activity against the human breast cancer SKBR-3 cell line. Gómez-Pérez V, McSorley T, See Too WC, Konrad M, Campos JM. ChemMedChem 7 663-669 (2012)
  4. Kinetic and mechanistic characterisation of Choline Kinase-α. Hudson CS, Knegtel RM, Brown K, Charlton PA, Pollard JR. Biochim. Biophys. Acta 1834 1107-1116 (2013)
  5. Phosphatidylcholine metabolism and choline kinase in human osteoblasts. Li Z, Wu G, van der Veen JN, Hermansson M, Vance DE. Biochim. Biophys. Acta 1841 859-867 (2014)
  6. Determination of potential scaffolds for human choline kinase α1 by chemical deconvolution studies. Sahún-Roncero M, Rubio-Ruíz B, Conejo-García A, Velázquez-Campoy A, Entrena A, Hurtado-Guerrero R. Chembiochem 14 1291-1295 (2013)
  7. New non-symmetrical choline kinase inhibitors. Schiaffino-Ortega S, López-Cara LC, Ríos-Marco P, Carrasco-Jimenez MP, Gallo MA, Espinosa A, Marco C, Entrena A. Bioorg. Med. Chem. 21 7146-7154 (2013)
  8. Antiplasmodial activity and mechanism of action of RSM-932A, a promising synergistic inhibitor of Plasmodium falciparum choline kinase. Zimmerman T, Moneriz C, Diez A, Bautista JM, Gómez Del Pulgar T, Cebrián A, Lacal JC. Antimicrob. Agents Chemother. 57 5878-5888 (2013)
  9. A new family of choline kinase inhibitors with antiproliferative and antitumor activity derived from natural products. Estévez-Braun A, Ravelo AG, Pérez-Sacau E, Lacal JC. Clin Transl Oncol 17 74-84 (2015)
  10. New more polar symmetrical bipyridinic compounds: new strategy for the inhibition of choline kinase α1. Castro-Navas FF, Schiaffino-Ortega S, Carrasco-Jimenez MP, Ríos-Marco P, Marco C, Espinosa A, Gallo MA, Mariotto E, Basso G, Viola G, Entrena-Guadix A, López-Cara LC. Future Med Chem 7 417-436 (2015)
  11. Pharmacophore-Based Virtual Screening to Discover New Active Compounds for Human Choline Kinase α1. Serrán-Aguilera L, Nuti R, López-Cara LC, Mezo MÁ, Macchiarulo A, Entrena A, Hurtado-Guerrero R. Mol Inform 34 458-466 (2015)
  12. Structural and enzymatic characterization of the choline kinase LicA from Streptococcus pneumoniae. Wang L, Jiang YL, Zhang JR, Zhou CZ, Chen Y. PLoS ONE 10 e0120467 (2015)
  13. Upregulation of Nicotinic Acetylcholine Receptor alph4+beta2 through a Ligand-Independent PI3Kbeta Mechanism That Is Enhanced by TNFalpha and the Jak2/p38Mapk Pathways. Rogers SW, Gahring LC. PLoS ONE 10 e0143319 (2015)
  14. Phosphorylation of Human Choline Kinase Beta by Protein Kinase A: Its Impact on Activity and Inhibition. Chang CC, Few LL, Konrad M, See Too WC. PLoS ONE 11 e0154702 (2016)
  15. Plasmodium falciparum Choline Kinase Inhibition Leads to a Major Decrease in Phosphatidylethanolamine Causing Parasite Death. Serrán-Aguilera L, Denton H, Rubio-Ruiz B, López-Gutiérrez B, Entrena A, Izquierdo L, Smith TK, Conejo-García A, Hurtado-Guerrero R. Sci Rep 6 33189 (2016)