The Protein-Protein Interaction Studies Between BMP-1 And BMP-4

Bone Morphogenetic Proteins(BMPs)are secreted molecules which play essential roles in early embryonic development.They are members of Transforming Growth Factor-?(TGF-?),which regulate cell proliferation and differentiation by binding to cell surface receptors.Some BMPs,such as BMP-4,are acting as morphogen to regulate the Dorsal-Ventral patterning in embryogenesis by establishing concentration-gradient in extracellular matrix.BMP-1 is actually a member of the Tolloid family of the astacin metalloproteinases.BMP-1 could regulate BMP signaling by liberating BMPs from complexes of BMP and its antagonist Chordin.The regulation network among these secreted molecules in extracellular matrix results in a stable BMP gradient along the D-V axis,controlling the cell fate of germ layers of embryo and neurogenesis.However,BMP could act as a negative regulator of BMP-1 to down-regulate the activity by binding its non-catalytic domain,and thus forms a negative feedback loop to limit BMP-1's activity.Our experiments aim on clarifying these interactions between BMP-1 and BMP-4 for understanding how BMP grandient formed.Herein,we constructed vectors for recombinant protein expression in the 293 E cell line.It is a high-throughput recombinant protein production system of suspesion-growing for secreted protein.We generated stable cell line for BMP-4,and sorted out the best monoclone of high-level expression.We also purified functional BMP-4 and tested its inducing activity through C2C12 cell line.We tried to purified BMP-1 for accumulating some experience,and more detailed work could start on purifying and functional analysing experiment in the future.