title: Protection of neuronal calcium sensor 1 protein in cells treated with paclitaxel.
abstract: Paclitaxel (Taxol) is one of the most effective treatment options for patients suffering from a variety of cancers. A major side effect seen in a high percentage of patients treated with paclitaxel is irreversible peripheral neuropathy. We previously reported that prolonged treatment with paclitaxel activates a calcium-dependent enzyme, calpain, which degrades neuronal calcium sensor 1 (NCS-1) and subsequent loss of intracellular calcium signaling. Because it appears that activation of calpain is an early step in this destructive cascade, we proposed that inhibition of calpain will protect against the unwanted side effects of paclitaxel treatment. First, NCS-1 levels and intracellular calcium signaling were found to be protected by the presence of lactacystin, a protesome inhibitor. To reinforce the role of calpain in this process, we showed that increased concentrations of calpastatin, a naturally occurring calpain inhibitor, were protective. Next, we tested two mutated versions of NCS-1 developed with point mutations at the P2 position of the calpain cleavage site of NCS-1 to decrease the likelihood of NCS-1 degradation. One mutant was cleaved more favorably by calpain compared with NCS-1 WT, whereas the other mutant was less favorably cleaved. Expression of either mutated version of NCS-1 in neuroblastoma cells protected intracellular calcium signals from paclitaxel-induced changes. These results support our hypothesis that it is possible to protect cells from paclitaxel-induced degradation of NCS-1 by inhibiting calpain activity.
authors.affiliation: Department of Pharmacology and Cellular and Molecular Physiology, Yale University, New Haven, Connecticut 06520, USA.
authors.forename: Jennifer H
authors.initials: JH
authors.lastname: Benbow
authors.forename: Brenda
authors.initials: B
authors.lastname: DeGray
authors.forename: Barbara E
authors.initials: BE
authors.lastname: Ehrlich
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D020013:Calcium Signaling; D002154:Calpain / Q000378:metabolism*; D045744:Cell Line, Tumor; D006801:Humans; D009154:Mutation; D009447:Neuroblastoma / Q000378:metabolism; D051580:Neuronal Calcium-Sensor Proteins / Q000187:drug effects / Q000378:metabolism*; D009474:Neurons / Q000473:pathology; D009479:Neuropeptides / Q000187:drug effects / Q000378:metabolism*; D017239:Paclitaxel / Q000494:pharmacology*; D017354:Point Mutation; D011480:Protease Inhibitors / Q000494:pharmacology; D020033:Protein Isoforms; D015398:Signal Transduction; D013816:Thermodynamics
chemical_list: D051580:Neuronal Calcium-Sensor Proteins; D009479:Neuropeptides; D011480:Protease Inhibitors; D020033:Protein Isoforms; C101464:frequenin calcium sensor proteins; C067713:lactacystin; D002154:Calpain; D017239:Paclitaxel; D000111:Acetylcysteine