1. Inactivation of the glutamine/amino acid transporter ASCT2 by 1,2,3-dithiazoles: proteoliposomes as a tool to gain insights in the molecular mechanism of action and of antitumor activity
- Author
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Oppedisano, F., Catto, M., Koutentis, Panayiotis Andreas, Nicolotti, O., Pochini, L., Koyioni, Maria G., Introcaso, A., Michaelidou, Sophia S., Carotti, A., Indiveri, C., Koutentis, Panayiotis Andreas [0000-0002-4652-7567], and Koyioni, Maria [0000-0002-2786-7523]
- Subjects
Amino Acid Transport Systems ,Glutamine ,Antiporter ,Toxicology ,amino acid transporter ,covalent bond ,Neoplasms ,dose response ,rat ,cysteine ,antineoplastic agent ,Cancer ,chemistry.chemical_classification ,Chemistry ,article ,amino acid transporter alanine serine cysteine transporter 2 ,ASCT2 ,unclassified drug ,Amino acid ,dithioerythritol ,Biochemistry ,1,2,3-dithiazoles ,computer analysis ,Amino Acid Transport System ASC ,Stereochemistry ,Proteolipids ,animal experiment ,Transport ,Antineoplastic Agents ,antineoplastic activity ,animal tissue ,Minor Histocompatibility Antigens ,Structure-Activity Relationship ,[(4 chloro 5 h 1,2,3 dithiazol 5 ylidene)amino]arenes ,proteoliposome ,inhibition kinetics ,drug mechanism ,Animals ,2 (4 chloro 5 h 1,2,3 dithiazol 5 ylidene)arene 1 (2 h) ones ,Structure–activity relationship ,Computer Simulation ,controlled study ,Cysteine ,Amino acid transporter ,Binding site ,enzyme substrate complex ,Pharmacology ,structural homology ,nonhuman ,Binding Sites ,Dose-Response Relationship, Drug ,Rattus ,molecular library ,IC 50 ,Transporter ,Rats ,Kinetics ,Thiazoles ,Liposomes ,thiol group - Abstract
The ASCT2 transport system catalyses a sodium-dependent antiport of glutamine and other neutral amino acids which is involved in amino acid metabolism. A library of 1,2,3-dithiazoles was designed, synthesized and evaluated as inhibitors of the glutamine/amino acid ASCT2 transporter in the model system of proteoliposomes reconstituted with the rat liver transporter. Fifteen of the tested compounds at concentration of 20μM or below, inhibited more than 50% the glutamine/glutamine antiport catalysed by the reconstituted transporter. These good inhibitors bear a phenyl ring with electron withdrawing substituents. The inhibition was reversed by 1,4-dithioerythritol indicating that the effect was likely owed to the formation of mixed sulfides with the protein's Cys residue(s). A dose-response analysis of the most active compounds gave IC50 values in the range of 3-30μM. Kinetic inhibition studies indicated a non-competitive inhibition, presumably because of a potential covalent interaction of the dithiazoles with cysteine thiol groups that are not located at the substrate binding site. Indeed, computational studies using a homology structural model of ASCT2 transporter, suggested as possible binding targets, Cys-207 or Cys-210, that belong to the CXXC motif of the protein. © 2012 Elsevier Inc. 265 1 93 102 Cited By :23
- Published
- 2012