Identification of cytosolic aldehyde dehydrogenase 1 from non-small cell lung carcinomas as a flavopiridol-binding protein.
The synthetic flavone flavopiridol can be cytostatic or cytotoxic to mammalian cells, depending on the concentration of the drug and the duration of exposure. It has been shown to inhibit the cyclin-dependent kinase (CDK) family of cell cycle regulatory enzymes. However, the existence of additional potential targets for drug action remains a matter of interest to define. To identify cellular targets, flavopiridol was immobilized. CDKs, particularly CDK 4, bound weakly to immobilized flavopiridol when ATP was absent but not in its presence. Two proteins with molecular weights of 40 kDa and 120 kDa had high affinities to the immobilized flavopiridol independent of the presence of ATP. They were present in all cell lines analyzed: cervical (HeLa), prostate and non-small cell lung carcinoma (NSCLC) cell lines. A 60-kDa protein, which was present only in NSCLC cells and bound similarly well to immobilized flavopiridol, was identified as cytosolic aldehyde dehydrogenase class 1 (ALDH-1). The level of this protein correlated with the resistance of NSCLC cell lines to cytotoxicity caused by 500 nM flavopiridol but not higher flavopiridol concentrations. Despite binding to ALDH-1, there was no inhibition of dehydrogenase activity by flavopiridol concentrations as high as 20 microM and flavopiridol was not metabolized by ALDH-1. The results suggest that high cellular levels of ALDH-1 may reduce cytotoxicity of flavopiridol and contribute to relative resistance to the drug. This is the first report that flavopiridol binds to proteins other than CDKs.
Schnier
J B
JB
Department of Biological Chemistry, University of California Davis, 95616, USA. jbschnier@ucdavis.edu
Kaur
G
G
Kaiser
A
A
Stinson
S F
SF
Sausville
E A
EA
Gardner
J
J
Nishi
K
K
Bradbury
E M
EM
Senderowicz
A M
AM
D000444:Aldehyde Dehydrogenase / Q000378:metabolism*; D000080924:Aldehyde Dehydrogenase 1 Family; D002289:Carcinoma, Non-Small-Cell Lung / Q000201:enzymology*; D002352:Carrier Proteins; D002846:Chromatography, Affinity; D003600:Cytosol / Q000201:enzymology*; D004305:Dose-Response Relationship, Drug; D005419:Flavonoids / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008175:Lung Neoplasms / Q000201:enzymology*; D010880:Piperidines / Q000378:metabolism*; D050697:Retinal Dehydrogenase; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay
D002352:Carrier Proteins; D005419:Flavonoids; D007527:Isoenzymes; D010880:Piperidines; C077990:alvocidib; D000080924:Aldehyde Dehydrogenase 1 Family; D000444:Aldehyde Dehydrogenase; C510223:ALDH1A1 protein, human; D050697:Retinal Dehydrogenase