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.
- © 1993 The Biochemical Society, London