Identification and characterization of a novel heparin-binding peptide for promoting osteoblast adhesion and proliferation by screening an Escherichia coli cell surface display peptide library.
Heparin/heparan sulfate (HS) plays a key role in cellular adhesion. In this study, we utilized a 12-mer random Escherichia coli cell surface display library to identify the sequence, which binds to heparin. Isolated insert analysis revealed a novel heparin-binding peptide sequence, VRRSKHGARKDR, designated as HBP12. Our analysis of the sequence alignment of heparin-binding motifs known as the Cardin-Weintraub consensus (BBXB, where B is a basic residue) indicates that the HBP12 peptide sequence contains two consecutive heparin-binding motifs (i.e. RRSK and RKDR). SPR-based BIAcore technology demonstrated that the HBP12 peptide binds to heparin with high affinity (KD = 191 nM). The HBP12 peptide is found to bind the cell surface HS expressed by osteoblastic MC3T3 cells and promote HS-dependent cell adhesion. Moreover, the surface-immobilized HBP12 peptide on titanium substrates shows significant increases in the osteoblastic MC3T3-E1 cell adhesion and proliferation.
Kim
Hyoun-Ee
HE
School of Materials Science and Engineering, Seoul National University, Seoul 151-742, Republic of Korea.
Kim
Hae-Won
HW
Jang
Jun-Hyeog
JH
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006493:Heparin / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism*; D019151:Peptide Library; D011485:Protein Binding
D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D019151:Peptide Library; D006493:Heparin