EXPRESSION AND CHARACTERIZATION OF HUMAN BONE MORPHOGENETIC PROTEIN-2 IN SILKWORM LARVAE INFECTED WITH RECOMBINANT BOMBYX-MORI NUCLEAR POLYHEDROSIS-VIRUS

被引:26
作者
ISHIDA, N
TSUJIMOTO, M
KANAYA, T
SHIMAMURA, A
TSURUOKA, N
KODAMA, S
KATAYAMA, T
OIKAWA, S
MATSUI, M
NAKANISHI, T
KOBAYASHI, J
NAKAZATO, K
机构
[1] KATAKURA IND,BIOL SCI RES INST,CHUOU KU,MATSUMOTO,NAGANO 390,JAPAN
[2] KATAKURA IND,CHUO SANKEN LAB,SAITAMA 35013,JAPAN
[3] NATL INST SERICULTURAL & ENTOMOL SCI,DEPT INSECT GENET & BREEDING,TSUKUBA,IBARAKI 305,JAPAN
关键词
BACULOVIRUS; BONE MORPHOGENETIC PROTEIN; BMNPV; GLYCOSYLATION; SILKWORM;
D O I
10.1093/oxfordjournals.jbchem.a124329
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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-E1 cells and ectopic bone formation in mice. On SDS-polyacrylamide gel electrophoretic analysis, the purified protein showed a 16kDa band under reducing conditions and a 30 kDa band under non-reducing conditions. The silkworm-expressed rhBMP-2 was glycosylated and susceptible to endo-beta-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.
引用
收藏
页码:279 / 285
页数:7
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