Characterization of the surface immobilized synthetic heparin binding domain derived from human fibroblast growth factor-2 and its effect on osteoblast differentiation.
Fibroblast growth factor (FGF)-2 regulates a variety of cellular functions, such as proliferation and differentiation, by binding to cell surface FGF receptors (FGFRs) in the presence of heparin proteoglycans. FGF-2 is known as a heparin-binding growth factor, but the localization of the heparin binding site has not been fully investigated until now. We used two potential heparin binding domains of FGF-2, the residues 105-111 (F105, YKRSRYT) and 119-135 (F119, KRTGQYKLGSKTGPGQK). Peptides could be stably immobilized onto the surface of tissue culture plates. Using solid phase binding assays, we demonstrated that both peptides had higher binding affinity toward heparin compared with nonbinding control sequence. The biological significance of these sites was tested by cell attachment and osteoblast differentiation studies. Cell attachment to the peptides F105 and F119 increased in a dose-dependent manner. Heparin and heparinase treatments decreased cell adhesion to both F105 and F119. This demonstrates that both F105 and F119 interact with cell-surface heparan sulfate proteoglycans, suggesting that FGF-2 has two heparin binding sites. In addition, osteoblast differentiation, confirmed by ALPase activity and mineralization, was increased by surface immobilized peptide F105 and F119. Taken together, these heparin binding peptides could be applied as biological agents enhancing osteoblast differentiation as well as surface modification tools in the tissue regeneration area, especially for bone regeneration.
Lee
Jue-Yeon
JY
Department of Craniomaxillofacial Reconstructive Science, School of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.|Intellectual Biointerface Engineering Center, College of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.
Choo
Jung-Eun
JE
Department of Craniomaxillofacial Reconstructive Science, School of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.|Intellectual Biointerface Engineering Center, College of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.
Choi
Young-Sook
YS
Department of Craniomaxillofacial Reconstructive Science, School of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.|Intellectual Biointerface Engineering Center, College of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.
Lee
Kuen-Yong
KY
Department of Bioengineering, College of Engineering, Hanyang University, 17 Haengdang-Dong, Seongdong-Gu, Seoul 133-791, South Korea.
Min
Do-Sik
DS
Department of Molecular Biology, College of Natural Sciences, Pusan National University, Geumjeong-Gu, Busan 609-735, South Korea.
Pi
Sung-Hee
SH
Department of Periodontology, College of Dentistry, WonKwang University, 344-2 Shinyong-Dong, Iksan, Chonbuk 570-749, South Korea.
Seol
Yang-Jo
YJ
Intellectual Biointerface Engineering Center, College of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.
Lee
Seung-Jin
SJ
Department of Pharmacy, College of Pharmacy, Ewha Womans University, 11-1 Daehyun-Dong, Seodaemun-Gu, Seoul 120-750, South Korea.
Jo
In-Ho
IH
Center for Biomedical Sciences, Korean National Institute of Health (KNIH), 5 Nokbun-Dong, Eunpyung-Gu, Seoul 122-701, South Korea.
Chung
Chong-Pyoung
CP
Intellectual Biointerface Engineering Center, College of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.|Department of Periodontology, School of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.
Park
Yoon-Jeong
YJ
Department of Craniomaxillofacial Reconstructive Science, School of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.|Intellectual Biointerface Engineering Center, College of Dentistry, Seoul National University, 28-22 Yongon-Dong, Chongno-Ku, Seoul 110-749, South Korea.
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D002454:Cell Differentiation; D002478:Cells, Cultured; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry* / Q000378:metabolism; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000166:cytology*; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction
D000199:Actins; D016222:Fibroblast Growth Factor 2; D006493:Heparin