title: Recombinant hBMP4 incorporated with non-canonical amino acid for binding to hydroxyapatite.
abstract: A novel growth factor containing non-canonical amino acids was designed and synthesized to enhance the binding to hydroxyapatite (HA). The designed protein was human bone morphogenetic protein 4 (hBMP4) incorporating diphosporylated serines (pSpS) that was found in salivary protein statherin and was reported to be responsible for binding to HA. Recombinant hBMP4 and a short peptide sequences containing pSpS were ligated by enzymatice reaction of sortase A, which exchanges the terminal amino acids of two polypeptides. Resulting hBMP4 containing pSpS (hBMP4-pSpS) bound HA more efficiently than hBMP-4 tagged with canonical serines (hBMP4-SS). The HA-bound hBMP-4-pSpS exhibited osteogenesis inducing activity to multipotential mesenchyme cells (C3H10T1/2) as evidenced by increased expression of osteogenic markers, which was not seen by hBMP4-SS. This novel protein with non-canonical serines will be applicable to bone regeneration materials in combination with HA.
authors.affiliation: Nano Medical Engineering Laboratory, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Sakuragi
authors.forename: Takashi
authors.initials: T
authors.lastname: Kitajima
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nagamune
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ito
mesh_terms: D019881:Aminoacyltransferases / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000378:metabolism; D017886:Durapatite / Q000378:metabolism*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000235:genetics* / Q000378:metabolism*
chemical_list: C525809:BMP4 protein, human; D001426:Bacterial Proteins; D055415:Bone Morphogenetic Protein 4; D011994:Recombinant Proteins; D012694:Serine; D017886:Durapatite; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases