ABSTRACT
During the last decade the key role of antimicrobial peptides in innate immunity has been argued. They were found in plants and in different phylogenic groups of animals (insects, amphibia, and even in mammals). We report the production of a human peptide antibiotic that was previously characterized as an EGF receptor tyrosine kinase inhibitor in epidermoid carcinoma A431/1522 cell subline overexpressing TGF alpha. It is a 3 kDa hydrophobic cationic peptide cytotoxic for different species of Gr+ and Gr- bacteria in micromolar concentration range, and demonstrating slight fungicidal activity.
Subject(s)
Anti-Bacterial Agents/pharmacology , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Protein Kinase Inhibitors , Transforming Growth Factor alpha/metabolism , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Carcinoma, Squamous Cell , Cell Line/chemistry , Cell Line/metabolism , Chromatography, High Pressure Liquid , Electrophoresis, Polyacrylamide Gel , HumansABSTRACT
Cells of human epidermoid carcinoma A431 were used for obtaining of the cell line constantly expressing TGF-alpha. Recombinant virions were obtained by introducing the proviral DNA into PA317 cells by means of electrotransfection. A protein with the EGF-competing activity was found in a conditioned media of the chosen clone A431/1522-4. The concentration of this protein was several times higher than in a conditioned media of wild type A431 cells. By means of electrophoresis, isoelectric focusing, and immunochemical analysis it was shown that the protein is TGF-alpha. Similarly to EGF, the extracted TGF-alpha entirely displaced 125I-EGF specifically bound to receptor. TGF-alpha produced by the A431/1522 cells also stimulated autophosphorylation of the EGF receptor.
