Biochemical properties of amphibian bone morphogenetic protein-4 expressed in CHO cells.
The biochemical properties of recombinant amphibian bone morphogenetic protein-4 (BMP-4), the cDNA of which has been cloned recently by screening of a Xenopus cDNA library, was characterized. The protein was expressed by the transfection of Chinese hamster ovary (CHO) cells with the cDNA cloned into expression vectors bearing a cytomegalovirus promoter or a simian virus 40 promoter. Northern-blot analysis showed that the latter vector was more efficient for Xenopus BMP-4 expression. Specific antiserum against Xenopus BMP-4 peptide demonstrated that the protein is synthesized as a large precursor, processed to the mature form and then secreted from the cells as a homodimer. Analysis of the biological activity in the conditioned medium revealed that Xenopus BMP-4 has a potent alkaline phosphatase-inducing activity on mouse osteoblastic cells.
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
A
A
Suzuki
S
S
Nishimatsu
A
A
Shoda
K
K
Takebayashi
K
K
Murakami
N
N
Ueno
D000818:Animals; D015153:Blotting, Western; D019485:Bone Morphogenetic Proteins; D016466:CHO Cells; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005822:Genetic Vectors; D006133:Growth Substances / Q000235:genetics; D046911:Macromolecular Substances; D015394:Molecular Structure; D008970:Molecular Weight; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014162:Transfection; D014982:Xenopus laevis
D019485:Bone Morphogenetic Proteins; D006133:Growth Substances; D046911:Macromolecular Substances; D011506:Proteins; D013762:Tetrahydrofolate Dehydrogenase