title: PDZ Domain in the Engineering and Production of a Saporin Chimeric Toxin as a Tool for targeting Cancer Cells.
abstract: In this paper we have studied a PDZ protein domain as a possible tool for cellular targeting of the ribosome inactivating protein Saporin, exploiting the ability of PDZ domains to recognize and bind short peptide sequences located at the C-terminus of a cognate protein. We have focused our attention on the PDZ domain from hCASK (Human calcium/calmodulin-dependent serine protein kinase) that binds extracellular CD98 in epithelial cells, being this antigen recognized as a marker for several human tumors and particularly considered a negative prognostic marker for human glioblastoma. We produced recombinant fusions of one or two hCASK-PDZ domains with the ribosome inactivating protein Saporin and assayed them on two human glioblastoma cell lines (GL15 and U87). These constructs proved to be toxic, with increasing activity as a function of the number of PDZ domains, and induce cell death by apoptotic mechanisms in a dose-dependent and/or time dependent manner.
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Giansanti
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.
authors.forename: Domenica
authors.initials: D
authors.lastname: Sabatini
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.
authors.forename: Maria Rosaria
authors.initials: MR
authors.lastname: Pennacchio
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Scotti
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Angelucci
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.
authors.forename: Anne-Chloè
authors.initials: AC
authors.lastname: Dhez
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Antonosante
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.|Sbarro Institute for Cancer Research and Molecular Medicine and Center for Biotechnology, Temple University, Philadelphia, Pennsylvania.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Cimini
authors.affiliation: Department of Medical, Surgery & Neurosciences, University of Siena, Italy.|Sbarro Institute for Cancer Research and Molecular Medicine and Center for Biotechnology, Temple University, Philadelphia, Pennsylvania.
authors.forename: Antonio
authors.initials: A
authors.lastname: Giordano
authors.affiliation: Department of Health, Life and Environmental Sciences, University of L'Aquila, I-67100 L'Aquila, Italy.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Ippoliti
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D027261:Fusion Regulatory Protein-1 / Q000737:chemistry / Q000378:metabolism*; D005909:Glioblastoma / Q000188:drug therapy / Q000276:immunology; D051528:Guanylate Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D000922:Immunotoxins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D058990:Molecular Targeted Therapy; D054731:PDZ Domains; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D054789:Ribosome Inactivating Proteins, Type 1 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000076985:Saporins
chemical_list: D027261:Fusion Regulatory Protein-1; D000922:Immunotoxins; D011994:Recombinant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; C106868:CASK kinases; D051528:Guanylate Kinases; D000076985:Saporins
keywords: CHIMERIC TOXINS; GLIOBLASTOMA; PDZ DOMAINS; SAPORIN; hCASK