Engineering of a multi-functional extracellular matrix protein for immobilization to bone mineral hydroxyapatite.
We have developed a strategy of designing multi-functional extracellular matrix proteins for functionalizing bone tissue engineering scaffolds and other biomedical surfaces to achieve improvements in bone grafting, bone repair and bone regeneration. We developed a novel extracellular matrix protein designed to have a cell adhesive RGD sequence derived from fibronectin and active functional unit of osteocalcin (OC) containing Ca(2+)-binding sites for immobilization to mineral component of bone, hydroxyapatite (HA). The fusion protein, designated FN(RGD)/OC, was expressed in Escherichia coli and purified with affinity chromatography using a His-tag. The resultant FN(RGD)/OC fusion protein preferentially bound to HA, promoted cell adhesive activity, and stimulated differentiation of MC3T3-E1 cell.
Kang
Wonmo
W
Department of Biochemistry, Inha University School of Medicine, Incheon, 400-712, Korea.
Kim
Tai-Il
TI
Yun
Yerang
Y
Kim
Hae-Won
HW
Jang
Jun-Hyeog
JH
D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D002846:Chromatography, Affinity; D017886:Durapatite / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010006:Osteoblasts / Q000502:physiology; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D011993:Recombinant Fusion Proteins; D015675:Osteocalcin; D017886:Durapatite