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PDZ Domain in the Engineering and Production of a Saporin Chimeric Toxin as a Tool for targeting Cancer Cells
Author(s) -
Giansanti Francesco,
Sabatini Domenica,
Pennacchio Maria Rosaria,
Scotti Stefano,
Angelucci Francesco,
Dhez AnneChloè,
Antonosante Andrea,
Cimini Annamaria,
Giordano Antonio,
Ippoliti Rodolfo
Publication year - 2015
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25080
Subject(s) - pdz domain , saporin , fusion protein , ribosome inactivating protein , microbiology and biotechnology , biology , recombinant dna , chemistry , biochemistry , cytotoxicity , ribosome , immunotoxin , in vitro , gene , rna
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. J. Cell. Biochem. 116: 1256–1266, 2015. © 2015 Wiley Periodicals, Inc.