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Modulation of choline kinase activity in human cancer cells observed by dynamic 31 P NMR
Author(s) -
Gabellieri Cristina,
BelouecheBabari Mounia,
Jamin Yann,
Payne Geoffrey S.,
Leach Martin O.,
Eykyn Thomas R.
Publication year - 2009
Publication title -
nmr in biomedicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.278
H-Index - 114
eISSN - 1099-1492
pISSN - 0952-3480
DOI - 10.1002/nbm.1361
Subject(s) - choline kinase , phosphocholine , choline , hela , enzyme , chemistry , kinase , protein kinase a , cancer cell , biochemistry , stereochemistry , cell , membrane , biology , cancer , phospholipid , genetics , phosphatidylcholine
Abstract Choline metabolites are widely studied in cancer research as biomarkers of malignancy and as indicators of therapeutic response. However, endogenous phosphocholine levels are determined by a number of processes that confound the interpretation of these measurements, including membrane transport rates and a series of enzyme catalysed reactions in the Kennedy pathway. Employing a dynamic 31 P NMR assay that is specific to choline kinase (ChoK) we have measured the rates of this enzyme reaction in cell lysates of MDA‐MB‐231 breast, PC‐3 prostate and HeLa cervical cancer cells and in solutions of purified human ChoK. The rates are sensitive to inhibition by hemicholinium‐3 (HC‐3), a competitive ChoK inhibitor, and to N ‐[2‐bromocinnamyl(amino)ethyl]‐5‐isoquinolinesulphonamide (H‐89), an agent commercialized as a specific cyclic‐AMP‐dependent protein kinase A (PKA) inhibitor. Copyright © 2009 John Wiley & Sons, Ltd.