Title : Expression and characterization of murine
osteoblast-specific factor 2 (
OSF-2 ) in a baculovirus expression system
Abstract :
- Osteoblast-specific factor 2 ( OSF-2 ) is a approximately 90-kDa protein selectively expressed in bone
- OSF-2 cDNA was recently isolated from mouse and human cDNA libraries and shows limited sequence homology with fasciclin I , a cell adhesion protein expressed in insect nerve cells
- Here we describe the expression of recombinant murine OSF-2 ( rmOSF-2 ) in a baculovirus/insect cell system
- Western blotting analysis employing polyclonal antiserum raised against a C-terminal synthetic OSF-2 peptide detected a protein of approximately 90-kDa as early as 2 days after infection of Sf9 cells with the recombinant virus
- Tunicamycin treatment of infected cells resulted in a mobility shift of OSF-2 (approximately 90-kDa band) on Western blots
- N-Glycanase digestion resulted in the same mobility shift of OSF-2 , indicating that rmOSF-2 expressed in insect cells is N-glycosylated
- However, OSF-2 was insensitive to endoglycosidase H digestion while a major fraction of this protein had affinity for concanavalin A. Finally, it was demonstrated that rmOSF-2 was able to bind to heparin
- This finding suggests that OSF-2 might be associated with the bone extracellular matrix after secretion by osteoblasts and participate in cell adhesion and/or cell communication
- The establishment of the baculovirus expression system with a high productivity of recombinant OSF-2 (around 40 micrograms/ml at maximum) and its heparin binding properties should allow us to obtain large amounts of rmOSF-2
Output (sent_index, trigger,
protein,
sugar,
site):
- 6. N-glycosylated, , rmOSF-2, -, -
Output(Part-Of) (sent_index,
protein,
site):
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):