Background: In our previous studies, we found that osteoactivin (OA) plays an important role in the regulation of osteoblast differentiation in vitro. Our studies also suggested that the region of OA protein that contains an RGD motif might play a vital role in the function of OA in osteoblast differentiation. In this study, we examined the functional role of OA-derived peptide containing the RGD motif (OA-D) in osteoblast differentiation.
Material/Methods: For this purpose, we designed another peptide, termed OA-E, that has sequence similar to OA-D but with glutamic acid (E) instead of aspartic acid (D). The effect of OA-E peptide on osteoblast differentiation was examined. Interestingly, OA-E peptide induced osteoblast differentiation in a manner similar to OA-D peptide. These data suggested that the effect of OA-derived peptides is RGD independent and it could be dependent on other features in the amino acid sequence of these peptides.
Results: OA-D peptide treatment markedly induced osteoblast differentiation markers in vitro compared to cultures treated with negative control peptide (NCP). Interestingly, OA-E peptide induced osteoblast differentiation in a manner similar to OA-D peptide. These data suggested that the effect of OA-derived peptides is RGD independent and it could be dependent on other features in the amino acid sequence of these peptides. Since phosphorylation of amino acid residues in proteins and peptides plays a major role in biological systems, the phosphorylation pattern of amino acid sequences of OA-derived peptides and OA protein family members were examined using bioinformatic analysis tools. We found that OA-derived peptides and OA protein family members have serine residue, close to c-terminus and might be phosphorylated with casein kinase II. Casein kinase II is known to phosphorylate many osteoblast-related proteins that regulate osteoblast development and differentiation such as osteopontin and vitronectin.
Conclusions: Collectively, these data showed that both OA-D and OA-E peptides significantly induced osteoblast differentiation in vitro and that effect is RGD independent.

Keywords: Amino Acid Sequence, Calcium - metabolism, Cell Line, Membrane Glycoproteins - physiology, Molecular Sequence Data, Oligopeptides - physiology, Osteoblasts - physiology, Osteocalcin - metabolism, Peptides - physiology