Author/Editor		Liović, Mirjana; Ozir, Mateja; Bedina-Zavec, Apolonija; Peternel, Špela; Komel, R; Zupančič, Tina
Title		Inclusion bodies as potential vehicles for recombinant protein delivery into epithelial cells
Type		članek
Source		Microb Cell Fact
Vol. and No.		Letnik 11, št. 1
Publication year		2012
Volume		str. 67
Language		eng
Abstract		Background: We present the potential of inclusion bodies (IBs) as a protein delivery method for polymeric filamentous proteins. We used as cell factory a strain of E. coli, a conventional host organism, and keratin 14 (K14) as an example of a complex protein. Keratins build the intermediate filament cytoskeleton of all epithelial cells. In order to build filaments, monomeric K14 needs first to dimerize with its binding partner (keratin 5, K5), which is then followed by heterodimer assembly into filaments. Results: K14 IBs were electroporated into SW13 cells grown in culture together with a reporter plasmid containing EYFP labeled keratin 5 (K5) cDNA. As SW13 cells do not normally express keratins, and keratin filaments are built exclusively of keratin heterodimers (i.e. K5/K14), the short filamentous structures we obtained in this study can only be the result of: a) if both IBs and plasmid DNA are transfected simultaneously into the cell(s); b) once inside the cells, K14 protein is being released from IBs; c) released K14 is functional, able to form heterodimers with EYFP-K5. Conclusions: Soluble IBs may be also developed for complex cytoskeletal proteins and used as nanoparticles for their delivery into epithelial cells.
Descriptors		RECOMBINANT PROTEINS INCLUSION BODIES DRUG DELIVERY SYSTEMS CYTOSKELETAL PROTEINS ELECTROPORATION TUMOR CELLS, CULTURED ESCHERICHIA COLI MICROSCOPY, ELECTRON, SCANNING