The Journal of Biochemistry
Online ISSN : 1756-2651
Print ISSN : 0021-924X
Expression and Characterization of Human Bone Morphogenetic Protein-2 in Silkworm Larvae Infected with Recombinant Bombyx mori Nuclear Polyhedrosis Virus
Nobuhiro IshidaMasafumi TsujimotoToshimichi KanayaAkio ShimamuraNobuo TsuruokaShiho KodamaToyoko KatayamaShinzo OikawaMasashi MatsuiToshihiro NakanishiJun KobayashiHiroshi Nakazato
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1994 Volume 115 Issue 2 Pages 279-285

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Abstract

Recombinant human bone morphogenetic protein-2 (rhBMP-2) was expressed in silkworm larvae, and a milligram quantity of the protein was purified and characterized. The expressed rhBMP-2 was biologically active in terms of induction of alkaline phosphatase activity in MC3T3-El cells and ectopic bone formation in mice. On SDS-polyacrylamide gel electrophoretic analysis, the purified protein showed a 16 kDa band under reducing conditions and a 30 kDa band under non-reducing conditions. The silkworm-expressed rhBMP-2 was glycosylated and susceptible to endo-β-N-acetylglucosaminidase F (endo F) and endo H, but resistant to endo D. Deglycosylated rhBMP-2 treated with endo F retained its biological activity. These results suggest that rhBMP-2 exists as a dimer and disulfide bond(s) are responsible for the dimerization. Moreover, sugar chains have no direct effect on the biological activity of the protein. The availability of a quite large amount of rhBMP-2 has allowed us to study the biological function of this interesting factor in detail.

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© The Japanese Biochemical Society
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