Cell adhesion biomaterial based on mussel adhesive protein fused with RGD peptide.
Previously, we designed and constructed a hybrid of the mussel adhesive protein (MAP) fp-151, which is a fusion protein with six type 1 (fp-1) decapeptide repeats at each type 5 (fp-5) terminus. Through various cell-adhesion analyses, we previously demonstrated that fp-151 has the potential to be used as a cell or tissue bioadhesive. In the present study, to improve the cell-adhesion properties of fp-151, we designed a new cell-adhesive protein, fp-151-RGD, which is a fusion with the GRGDSP residues, a RGD peptide sequence that has previously been identified at the cell-attachment site of fibronectin, at the C-terminus of fp-151. Although recombinant fp-151-RGD maintained the advantages associated with fp-151, such as a high production yield in Escherichia coli and simple purification, it showed superior spreading ability, which is important for cell proliferation under serum-free conditions, as well as better cell-adhesion ability compared with other commercially produced cell-adhesion materials such as poly-l-lysine (PLL) and the naturally extracted MAP mixture Cell-Tak. The excellent adhesion and spreading abilities of fp-151-RGD might be due to the fact that it utilizes three types of cell-binding mechanisms: DOPA adhesion of Cell-Tak, cationic binding force of PLL, and RGD sequence-mediated adhesion of fibronectin. Therefore, the new recombinant fp-151-RGD is suitable for use as a cell-adhesion material in cell culture or tissue engineering, and in any other area where efficient cell adhesion is required.
Hwang
Dong Soo
DS
Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea.
Sim
Sung Bo
SB
Cha
Hyung Joon
HJ
D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion / Q000502:physiology*; D049109:Cell Proliferation; D020099:Coated Materials, Biocompatible / Q000737:chemistry / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D004295:Dihydroxyphenylalanine / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008422:Materials Testing; D018625:Microscopy, Atomic Force; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013499:Surface Properties; D014443:Tyrosine / Q000378:metabolism
D020099:Coated Materials, Biocompatible; D009842:Oligopeptides; D011506:Proteins; D011993:Recombinant Fusion Proteins; C037403:adhesive protein, mussel; D014443:Tyrosine; D004295:Dihydroxyphenylalanine; C047981:arginyl-glycyl-aspartic acid