Abstract
In the current study, we designed four cyclic peptide analogues by incorporating two cysteine residues in a BMP-2 linear knuckle epitope in such a way that the active region of the peptide could be either inside or outside the cyclic ring. Bone morphogenetic protein receptor BMPRII was immobilized on the chip surface, and the interaction of the linear and cyclic peptide analogues was studied using surface plasmon resonance (SPR). From the affinity data, the peptides with an active region inside the cyclic ring had a higher binding affinity in comparison to the other peptides. To confirm that our affinity data are in line in vitro, we studied the expression levels of RUNX2 (runt-related transcription factor) and conducted an osteogenic marker alkaline phosphatase (ALP) assay and staining. Based on the affinity data and the in vitro experiments, peptide P-05 could be a suitable candidate for osteogenesis, with higher binding affinity and increased RUNX2 and ALP expression in comparison to the linear peptides.
Highlights
Growth factors play a major role in the wound healing process by recruiting different cell types at the site of injury and reducing the time for healing by enhancing the cell signaling [1,2]
Bone grafts are used for the recovery of a fractured bone, which takes a longer time for recovery, whereas bone grafts combined with Bone morphogenetic proteins (BMPs)-2 protein require less time [9]
It is known that the bone morphogenetic protein 2 (BMP-2) knuckle epitope peptide binds to the Bone morphogenetic protein receptor-II (BMPRII) receptor, so we studied the interaction of the peptides with BMPRII using surface plasmon resonance (SPR)
Summary
Growth factors play a major role in the wound healing process by recruiting different cell types (chemotaxis) at the site of injury and reducing the time for healing by enhancing the cell signaling [1,2]. In 2008, the FDA issued an alert that the usage of BMP-2 protein in spinal fusion in certain cases can lead to swelling of the neck and throat tissue, resulting in compression of the airway and/or the neurological structures in the neck [11]. In 2003, Saito et al fragmented the BMP-2 protein from the knuckle epitope region into seven small peptides, and all of these peptides (i.e., P1, P2, P3, P4, P4sp, P5, and P6) were screened for alkaline phosphatase (ALP) activity [18]. C2C12 cells, upon treatment with BMP-2 protein, convert the myogenic pathway to the osteogenic pathway [24]
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