Gudivada Vijaya Narasimha, Huang Chen-Ji, Luo Yueh-Hsia, Dong Guo-Chung
Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 350, Taiwan.
Department of Life Sciences, National Central University, Taoyuan City 32001, Taiwan.
Polymers (Basel). 2021 Jul 31;13(15):2549. doi: 10.3390/polym13152549.
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.
在当前研究中，我们通过在骨形态发生蛋白-2（BMP-2）线性关节表位中引入两个半胱氨酸残基来设计四种环肽类似物，使得肽的活性区域可以位于环内或环外。将骨形态发生蛋白受体BMPRII固定在芯片表面，并使用表面等离子体共振（SPR）研究线性和环肽类似物的相互作用。根据亲和力数据，与其他肽相比，活性区域位于环内的肽具有更高的结合亲和力。为了在体外证实我们的亲和力数据是一致的，我们研究了RUNX2（ runt相关转录因子）的表达水平，并进行了成骨标志物碱性磷酸酶（ALP）测定和染色。基于亲和力数据和体外实验，与线性肽相比，肽P-05具有更高的结合亲和力以及更高的RUNX2和ALP表达，可能是成骨的合适候选物。
