Construction of Chimeric Human Epidermal Growth Factor Containing Short Collagen-Binding Domain Moieties for Use as a Wound Tissue Healing Agent

Abstract

Among the various human growth factors, epidermal growth factor (hEGF, consisting of 53 amino acids) has various effects on cell regeneration, stimulation of proliferation, migration of keratinocytes, formation of granulation tissues, and stimulation of fibroblast motility, which are important for wound healing. Owing to their multiple activities, EGFs are used as pharmaceutical and cosmetic agents. However, their low productivity, limited target specificity, and short half-life inhibit their application as therapeutic agents. To overcome these obstacles, we fused the collagen-binding domain (CBD) of Vibrio mimicus metalloprotease to EGF protein. About 18 or 12 amino acids (aa) (of the 33 total amino acids), which were essential for collagen-binding activity, were combined with the N- and C-termini of EGF. We constructed, expressed, and purified EGF (53 aa)-CBD (18 aa), EGF (53 aa)-CBD (12 aa), CBD (18 aa)-EGF (53 aa), and CBD (12 aa)-EGF (53 aa). These purified recombinant proteins increased the numbers of cells in treated specimens compared with non-treated specimens and control hEGF samples. The collagen-binding activities were also evaluated. Furthermore, CBD-hybridized hEGF induced phosphorylation of the EGF receptor. These results suggested that these fusion proteins could be applicable as small therapeutic agents in wound tissue healing.

Targeted Delivery of 111In Into the Nuclei of EGFR Overexpressing Cells via Modular Nanotransporters With Anti-EGFR Affibody

“…MNT C-EGF and MNT N-EGF presented with different effects on MCF-7 proliferation depending on the position of the EGFligand module. Similar effects were reported for other EGFcontaining chimeric constructs (Kim et al, 2015). МNТ N-EGF may interact with EGFR like an epigen ligand whose C-terminal sequence is long compared with that of EGF.…”

“…When EGF is used as a ligand in drug delivery vehicles, it can affect cellular signaling and stimulates events leading to enhances cancer cell proliferation (Chen et al, 2002;Kim et al, 2015). EGF and other natural ligands induce dimerization and autophosphorylation of EGFR (Schlessinger, 2002) and subsequent internalization (Friedman and Stahl, 2009) of the receptor.…”

Angiogenic Properties of ‘Leukocyte- and Platelet-Rich Fibrin’

“…reported high levels of EGF (513 pg/mL) in human platelet rich plasma (PRP) 16 . The low levels detected in our study are probably due to the short half-life and the rapid cell diffusion of EGF 44 . Unfortunately, in vitro studies cannot account for the influence of the physiological environment on the behavior of the platelet concentrate with regard to cellular crosstalk and growth factor release 32 .…”

C-terminal domains of bacterial proteases: structure, function and the biotechnological applications

“…Similarly, for the specific collagen‐binding activity of CBD, Kim discovered that human epidermal growth factor containing CBD could be used as a healing agent for wound tissue (Kim et al . ).…”