Rational Derivation of Osteogenic Peptides from Bone
Morphogenetic Protein-2 Knuckle Epitope by Integrating In
Silico Analysis and In Vitro Assay

Zhang, Aiying; Chen, Zhixia; Yu, Xiuli; Zhang, Li; Song, Qiyuan

doi:10.1007/s10989-020-10058-y

Cited by 6 publications

(5 citation statements)

References 46 publications

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“…Traditionally, it is thought that the knuckle epitope locates at the β 6 ‐ and β 7 ‐strand of BMP2 and covers a linear KL 73–92 region. However, it was found that not all residues in the KL 73–92 region can contribute effectively to the epitope interaction with BRII 17 . Therefore, we herein performed a systematic alanine scanning (SAS) 34 to derive the residue importance profile of BMP2 monomer in BMP2–BRII interaction.…”

Section: Resultsmentioning

confidence: 99%

“…the BMP2 binding domain of BRII receptor protein) was expressed in Baculovirus infected Sf9 cells and obtained commercially. The binding affinity of knuckle‐derived peptides to BRII was determined using a fluorescence spectroscopy protocol modified from our previous works 16,17 . Structural analysis revealed that the receptor binding pocket has two tryptophan residues (Trp45 and Trp60), which were used as fluorescent probes to detect the intrinsic tryptophan fluorescence change (Δ F ) upon peptide binding.…”

Section: Methodsmentioning

confidence: 99%

“…Previously, we have addressed considerable efforts on the rational design of linear and cyclic peptide ligands to specifically target the intermolecular interactions of BMP2 with its BRI and BRII receptors 16,17 . Recently, our co‐workers also described a new epitope termed armpit that represents the antagonist‐recognition site of BMP2 and is not effectively overlapped with the traditional wrist and knuckle epitopes 18 .…”

Section: Introductionmentioning

confidence: 99%

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Revisiting bone morphogenetic protein‐2 knuckle epitope and redesigning the epitope‐derived peptides

Zhao

Zhang

Che

et al. 2021

Journal of Peptide Science

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The bone morphogenetic protein‐2 (BMP2) plays a crucial role in bone formation, growth and regeneration, which adopts a conformational wrist epitope and a linear knuckle epitope to interact with its type‐I (BRI) and type‐II (BRII) receptors, respectively. In this study, we systematically examine the BRII‐recognition site of BMP2 at structural, energetic and dynamic levels and accurately locate hotspots of the recognition at BMP2–BRII complex interface. It is revealed that the traditional knuckle epitope (BMP2 residue range 73–92) do fully match the identified hotspots; the BMP2‐recognition site includes the C‐terminal region of traditional knuckle epitope as well as its flanked β‐strands. In addition, the protein context of full‐length BMP2 is also responsible for the recognition by addressing conformational constraint on the native epitope segment. Therefore, we herein redefine the knuckle epitope to BMP2 residue range 84–102, which has a similar sequence length but is slid along the protein sequence by ~10 residues as compared to traditional knuckle epitope. The redefined one is also a linear epitope that is natively a double‐stranded β‐sheet with two asymmetric arms as compared to the natively single β‐strand of the traditional version, although their sequences are partially overlapped to each other. It is revealed that the redefined epitope‐derived peptide LN84–102 exhibits an improved affinity by >3‐fold relative to the traditional epitope‐derived peptide KL73–92. Even so, the LN84–102 peptide still cannot fully represent the BMP2 recognition event by BRII that has been reported to have a nanomolar affinity. We further introduce a disulfide bond across the two arms of double‐stranded β‐sheet to constrain the free LN84–102 peptide conformation, which mimics the conformational constraint addressed by protein context. Consequently, several cyclic peptides are redesigned, in which the LN84–102(cyc89‐101) is determined to exhibit a sub‐micromolar affinity; this value is ~5‐fold higher than its linear counterpart. Structural analysis also reveals that the cyclic peptide can interact with BRII in a similar binding mode with the redefined knuckle epitope region in full‐length BMP2 protein.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Revisiting bone morphogenetic protein‐2 knuckle epitope and redesigning the epitope‐derived peptides

Zhao

Zhang

Che

et al. 2021

Journal of Peptide Science

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“…The P4 peptide comprises a modified sequence, KIPKASSVPTELSAISTLYL, corresponding to residues 73-92 of the knuckle epitope of bone morphogenetic protein 2 (BMP2), which is part of the BMP-binding site of BMP receptor II (BMPRII) [7][8][9]. The knuckle epitope of BMP 2 binds to the extracellular domain of BMPR II, and then the intracellular domain of BMPR II phosphorylates the type I receptor, which induces intracellular SMAD signaling [10].…”

Section: Introductionmentioning

confidence: 99%

“…BMP/SMAD signaling plays a critical role in the formation and maintenance of bone, cartilage and other tissues [11]. This P4 peptide has already been demonstrated to enhance bone formation, and it is believed to be due to the mechanisms above [7][8][9]12].…”

Section: Introductionmentioning

confidence: 99%

Biomimetic Silica Particles with Self-Loading BMP-2 Knuckle Epitope Peptide and Its Delivery for Bone Regeneration

Nguyen

Park

et al. 2023

Pharmaceutics

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Biomimetic silica deposition is an in-situ immobilization method for bioactive molecules under biocompatible conditions. The osteoinductive P4 peptide derived from the knuckle epitope of bone morphogenetic protein (BMP), which binds to BMP receptor-II (BMPRII), has been newly found to contain silica formation ability. We found that the two lysine residues at the N-terminus of P4 played a vital role in silica deposition. The P4 peptide co-precipitated with silica during P4-mediated silicification, yielding P4/silica hybrid particles (P4@Si) with a high loading efficiency of 87%. P4 was released from P4@Si at a constant rate for over 250 h, representing a zero-order kinetic model. In flow cytometric analysis, P4@Si showed a 1.5-fold increase in the delivery capacity to MC3T3 E1 cells than the free form of P4. Furthermore, P4 was found anchored to hydroxyapatite (HA) through a hexa-glutamate tag, followed by P4-mediated silicification, yielding P4@Si coated HA. This suggested a superior osteoinductive potential compared to silica or P4 alone coated HA in the in vitro study. In conclusion, the co-delivery of the osteoinductive P4 peptide and silica by P4-mediated silica deposition is an efficient method for capturing and delivering its molecules and inducing synergistic osteogenesis.

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Integrated In Silico-In Vitro Identification and Optimization of Bone Morphogenic Protein-2 Armpit Epitope as Its Antagonist Binding Site

Jia

Chi

et al. 2020

Protein J

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Rational Derivation of Osteogenic Peptides from Bone Morphogenetic Protein-2 Knuckle Epitope by Integrating In Silico Analysis and In Vitro Assay

Cited by 6 publications

References 46 publications

Revisiting bone morphogenetic protein‐2 knuckle epitope and redesigning the epitope‐derived peptides

Revisiting bone morphogenetic protein‐2 knuckle epitope and redesigning the epitope‐derived peptides

Biomimetic Silica Particles with Self-Loading BMP-2 Knuckle Epitope Peptide and Its Delivery for Bone Regeneration

Integrated In Silico-In Vitro Identification and Optimization of Bone Morphogenic Protein-2 Armpit Epitope as Its Antagonist Binding Site

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