Recombinant mussel adhesive protein Mgfp-5 as cell adhesion biomaterial.
Mytilus galloprovincialis foot protein type-5 (Mgfp-5) is one of the mussel adhesive proteins that participate in adhesion with the substratum. We previously reported the production of recombinant Mgfp-5 in Escherichia coli and showed that the recombinant protein had superior adhesion abilities versus those of Cell-Tak, a commercially available mussel adhesive protein mixture. In the present work, we investigated the feasibility of using recombinant Mgfp-5 as a cell adhesion agent. Purified and tyrosinase-modified recombinant Mgfp-5 was used to adhere living anchorage-independent cells such as insect Drosophila S2 cells and human MOLT-4 cells onto glass slides. Our results revealed that these cell lines efficiently attached to recombinant Mgfp-5-coated glass surfaces, and that surface-immobilized S2 cells were viable and able to undergo cell division for up to 1 week. Cytochemical studies with 4',6-diamidino-2-phenylindole (DAPI) staining of nuclei and immunofluorescence for secreted foreign human erythropoietin (hEPO) from recombinant S2 cells and quantitative comparative analyses of S2 cell binding ability with Cell-Tak and poly-L-lysine, the main cell adhesion agent, were performed to demonstrate successful usage of recombinant Mgfp-5 for cell biological applications. Collectively, these results indicate that recombinant Mgfp-5 may be a useful new cell adhesion biomaterial for anchorage-independent cells.
Hwang
Dong Soo
DS
Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea.
Gim
Youngsoo
Y
Kang
Dong Gyun
DG
Kim
Yeon Kyu
YK
Cha
Hyung Joon
HJ
D000818:Animals; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D004330:Drosophila / Q000166:cytology; D005898:Glass / Q000737:chemistry; D006801:Humans; D014442:Monophenol Monooxygenase / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013499:Surface Properties
D015815:Cell Adhesion Molecules; C518178:Mgfp-5 protein, Mytilus galloprovincialis; D011994:Recombinant Proteins; D014442:Monophenol Monooxygenase