Cdc25 inhibition and cell cycle arrest by a synthetic thioalkyl vitamin K analogue.
A synthetic vitamin K analogue, 2-(2-mercaptoethanol)-3-methyl-1,4-naphthoquinone or compound 5 (Cpd 5), was found previously to be a potent inhibitor of tumor cell growth. We now demonstrate that Cpd 5 arrested cell cycle progression at both G1 and G2-M. Because of the potential arylating activity of Cpd 5, it might inhibit Cdc25 phosphatases, which contain a cysteine in the catalytic site. To test this hypothesis, we examined the inhibitory activity of Cpd 5 against several cell cycle-relevant protein tyrosine phosphatases and found that Cpd 5 was a potent, selective, and partially competitive inhibitor of Cdc25 phosphatases. Furthermore, Cpd 5 caused time-dependent, irreversible enzyme inhibition, consistent with arylation of the catalytic cysteine in Cdc25. Treatment of cells with Cpd 5 blocked dephosphorylation of the Cdc25C substrate, Cdc2, and its kinase activity. Cpd 5 enhanced tyrosine phosphorylation of both potent regulators of G1 transition, ie., Cdk2 and Cdk4, and decreased the phosphorylation of Rb, an endogenous substrate for Cdk4 kinase. Furthermore, close chemical analogues that lacked in vitro Cdc25 inhibitory activity failed to block cell cycle progression and Cdc2 kinase activity. Cpd 5 did not alter the levels of p53 or the endogenous cyclin-dependent kinase inhibitors, p21 and p16. Our results support the hypothesis that the disruption in cell cycle transition caused by Cpd 5 was attributable to intracellular Cdc25 inhibition. This novel thioalkyl K vitamin analogue could be useful for cell cycle control studies and may provide a valuable pharmacophore for the design of future therapeutics.
Tamura
K
K
Department of Pharmacology, University of Pittsburgh, Pennsylvania 15261, USA.
Southwick
E C
EC
Kerns
J
J
Rosi
K
K
Carr
B I
BI
Wilcox
C
C
Lazo
J S
JS
D002453:Cell Cycle / Q000187:drug effects* / Q000502:physiology*; D006131:Growth Inhibitors / Q000494:pharmacology*; D006801:Humans; D008623:Mercaptoethanol / Q000031:analogs & derivatives* / Q000494:pharmacology; D009285:Naphthoquinones / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D014812:Vitamin K / Q000031:analogs & derivatives / Q000494:pharmacology; D020687:cdc25 Phosphatases / Q000502:physiology*
C111898:2-(2-mercaptoethanol)-3-methyl-1,4-naphthoquinone; D006131:Growth Inhibitors; D009285:Naphthoquinones; D014812:Vitamin K; D008623:Mercaptoethanol; D020687:cdc25 Phosphatases