Genetic grafting of membrane-acting peptides to the cytotoxin dianthin augments its ability to de-stabilize lipid bilayers and enhances its cytotoxic potential as the component of transferrin-toxin conjugates
| Autor: | Laura Costantini, Marco Colombatti, Giuseppe Tridente, Giulio Fracasso, G. Menestrina, E. Chiesa, Cristina Potrich, Irene Lorenzetti, Alessandra Meneguzzi, Giuseppe Legname |
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| Jazyk: | angličtina |
| Rok vydání: | 2000 |
| Předmět: |
Cancer Research
Recombinant Fusion Proteins Lipid Bilayers Ribosome Inactivating Proteins Hemagglutinin (influenza) Peptide law.invention Jurkat Cells Immunotoxin law Humans Monensin Lipid bilayer Plant Proteins chemistry.chemical_classification Membranes biology Immunotoxins Transferrin Membranes Artificial Hydrogen-Ion Concentration biology.organism_classification Fusion protein Oncology chemistry Biochemistry Vesicular stomatitis virus Artificial Type 1 biology.protein Recombinant DNA Ribosome Inactivating Proteins Type 1 Protein G |
| Popis: | Three chimeric proteins were obtained by fusing together the dianthin gene and DNA fragments encoding for the following membrane-acting peptides: the N-terminus of protein G of the vesicular stomatitis virus (KFT25), the N terminus of the HA2 hemagglutinin of influenza virus (pHA2), and a membrane-acting peptide (pJVE). Chimeric dianthins (KFT25DIA, pHA2DIA and pJVEDIA) retained full enzymatic activity in cell-free assays and showed increased ability to induce pH-dependent calcein release from large unilamellar vesicles (LUVs). pHA2DIA and pJVEDIA also showed faster kinetics of interaction with LUVs, while KFT25DIA and pHA2DIA displayed a reduced cytotoxicity as compared to wild-type dianthin. Conjugates made by chemically cross-linking KFT25DIA or pJVEDIA and human transferrin (Tfn) showed greater cell-killing efficiency than conjugates of Tfn and wild-type dianthin. As a consequence, by fusion of membrane-acting peptides to the dianthin sequence the specificity factor (i.e., the ratio between non-specific and specific toxicity) of Tfn-KFT25DIA, Tfn-pHA2DIA and Tfn-pJVEDIA was increased with respect to that of Tfn-based conjugates made with wild-type dianthin. Taken together, our results suggest that genetic fusion of membrane-acting peptides to enzymatic cytotoxins results in the acquisition of new physico-chemical properties exploitable for designing new recombinant cytotoxins and to tackle cell-intoxication mechanisms. Int. J. Cancer 86:582–589, 2000. © 2000 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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