Enhancement of sphingosine kinase 1 catalytic activity by deletion of 21 amino acids from the COOH-terminus.

Sphingosine kinase 1 (SphK1) responds to a variety of growth factor signals by increasing catalytic activity as it translocates to the plasma membrane (PM). Several studies have identified amino acids residues involved in translocation yet how SphK1 increases its catalytic activity remains to be elucidated. Herein, we report that deletion of 21 amino acids from the COOH-terminus of SphK1 (1-363) results in increased catalytic activity relative to wild-type SphK1 (1-384) which is independent of the phosphorylation state of Serine 225 and PMA stimulation. Importantly, HEK293 cells stably expressing the 1-363 protein exhibit enhanced cell growth under serum-deprived cell culture conditions. Together the evidence indicates that the COOH-terminal region of SphK1 encompasses a structural element that is necessary for the increase in catalytic activity in response to PMA treatment and that its deletion renders SphK1 constitutively active with respect to PMA treatment.

Sphingosine kinase 1 localized to the plasma membrane lipid raft microdomain overcomes serum deprivation induced growth inhibition.

Several studies have demonstrated that sphingosine kinase 1 (SphK1) translocates to the plasma membrane (PM) upon its activation and further suggested the plasma membrane lipid raft microdomain (PMLRM) as a target for SphK1 relocalization. To date, however, direct evidence of SphK1 localization to the PMLRM has been lacking. In this report, using multiple biochemical and subcellular fractionation techniques we demonstrate that endogenous SphK1 protein and its substrate, D-erythro-sphingosine, are present within the PMLRM. Additionally, we demonstrate that the PMA stimulation of SphK1 localized to the PMLRM results in production of sphingosine-1-phosphate as well as induction of cell growth under serum deprivation conditions. We further report that Ser225Ala and Thr54Cys mutations, reported to abrogate phosphatidylserine binding, block SphK1 targeting to the PMLRM and SphK1 induced cell growth. Together these findings provide direct evidence that the PMLRM is the major site of action for SphK1 to overcome serum-deprived cell growth inhibition.

Natural products as inhibitors of carcinogenesis.

BACKGROUND: Although carcinogenesis and cancer have been studied intensively for > 50 years, the rates of cancer incidence and mortality remain high. The most successful approach for reducing these rates has been primary prevention. For individuals at a high risk of developing cancer owing to certain genetic, environmental and occupational factors, cancer chemoprevention is a logical approach. OBJECTIVE: This review discusses natural products as inhibitors of carcinogenesis. CONCLUSION: Natural product chemopreventive agents are inherently biologically active and have been demonstrated to prevent and reverse the carcinogenic process in a pleiotropic manner. Derivatives of compounds discovered by screening natural products for chemopreventive agents have shown efficacy in clinical trials. Despite the obstacles that must be overcome before chemopreventive drugs become an integral component of standard medical practice for cancer prophylaxis, there is vast potential for significant improvement in cancer morbidity and mortality through the use of natural product chemopreventive drugs.