title: Fusion with pep-1, a cell-penetrating peptide, enhances the transmembrane ability of human epidermal growth factor.
abstract: Administration of macromolecule compositions in medicine and cosmetics always exhibited low bioavailability due to the limitation of transmembrane transport. Here, human epidermal growth factor (hEGF) was fused with glutathione S-transferase (GST) and Pep-1, the first commercial cell-penetrating peptide, in Escherichia coli. The fusion protein was firstly purified with the affinity chromatography, and then the GST tag was released by TEV protease. Final purification was achieved by the ion exchange chromatography. The biological activities and the transmembrane ability of the obtained products were determined using scratch wound-healing assay, MTT analysis, and immunofluorescence assay. The results showed that both rhEGF and Pep-1-fused hEGF were soluble expressed in E. coli. The fusion of Pep-1 could markedly increase the transmembrane ability of EGF, whereas it did not interfere with the growth-stimulating and migration-promoting functions of hEGF on fibroblasts. This research provided a novel strategy for the transmembrane transport of protein-derived cosmetics or drugs.
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Xue-Gang
authors.initials: XG
authors.lastname: Luo
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: De-Yun
authors.initials: DY
authors.lastname: Ma
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Wen
authors.initials: W
authors.lastname: Li
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Chong-Xi
authors.initials: CX
authors.lastname: Wang
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Ying-Ying
authors.initials: YY
authors.lastname: He
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Xiang-Chao
authors.initials: XC
authors.lastname: Gu
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Xiu-Mei
authors.initials: XM
authors.lastname: Li
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Hao
authors.initials: H
authors.lastname: Zhou
authors.affiliation: a Key Lab of Industrial Fermentation Microbiology (Tianjin University of Science and Technology) , Ministry of Education , Tianjin , P.R. China.|b Tianjin Key Lab of Industrial Microbiology , College of Biotechnology, Tianjin University of Science and Technology , Tianjin , P.R. China.
authors.forename: Tong-Cun
authors.initials: TC
authors.lastname: Zhang
mesh_terms: D000818:Animals; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism*; D002522:Chlorocebus aethiops; D002852:Chromatography, Ion Exchange; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000737:chemistry; D004815:Epidermal Growth Factor / Q000378:metabolism*; D006801:Humans; D010455:Peptides / Q000737:chemistry*
chemical_list: C503924:Pep-1 peptide; D010455:Peptides; D003543:Cysteamine; D004815:Epidermal Growth Factor
keywords: E. coli; EGF; Pep-1; expression; transmembrane