In silico design of fusion keratinocyte growth factor containing collagen-binding domain for tissue engineering application

Amidzadeh, Zahra; Rismani, Elham; Shokrgozar, Mohammad Ali; Rahimi, Hamzeh; Golkar, Majid

doi:10.1016/j.jmgm.2022.108351

Cited by 4 publications

(4 citation statements)

References 67 publications

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“…Amidzadeh et al designed a novel fusion protein that consists of KGF (keratinocyte growth factor) and cCBD, and their study revealed that the secondary structure of the recombinant peptide was better preserved when cCBD was fused to the C-terminal of KGF. 59 A possible explanation for these results is that the cCBD is a hydrophilic peptide, so this hydrophilic tail contributes to the conformational stability by forming a hydrogen bond with water. 60−62 Our earlier findings reasoned that the electrostatic interactions are likely involved in the binding of LL37 and cCBD-LL37 to collagen.…”

Section: Discussionmentioning

confidence: 99%

“…Although LL37 and cCBD-LL37 showed similar helicity changing behavior under varying pH, cCBD-LL37 exhibited less helicity changes. Amidzadeh et al designed a novel fusion protein that consists of KGF (keratinocyte growth factor) and cCBD, and their study revealed that the secondary structure of the recombinant peptide was better preserved when cCBD was fused to the C-terminal of KGF . A possible explanation for these results is that the cCBD is a hydrophilic peptide, so this hydrophilic tail contributes to the conformational stability by forming a hydrogen bond with water. − …”

Section: Discussionmentioning

confidence: 99%

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Characterization of LL37 Binding to Collagen through Peptide Modification with a Collagen-Binding Domain

Wei,

Rolle,

Camesano

2023

ACS Omega

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Collagen-based biomaterials loaded with antimicrobial peptides (AMPs) present a promising approach for promoting wound healing while providing protection against infections. In our previous work, we modified the AMP LL37 by incorporating a collagen-binding domain (cCBD) as an anchoring unit for collagen-based wound dressings. We demonstrated that cCBD-modified LL37 (cCBD-LL37) exhibited improved retention on collagen after washing with PBS. However, the binding mechanism of cCBD-LL37 to collagen remained to be elucidated. In this study, we found that cCBD-LL37 showed a slightly higher affinity for collagen compared to LL37. Our results indicated that cCBD inhibited cCBD-LL37 binding to collagen but did not fully eliminate the binding. This suggests that cCBD-LL37 binding to collagen may involve more than just one-site-specific binding through the collagen-binding domain, with non-specific interactions also playing a role. Electrostatic studies revealed that both LL37 and cCBD-LL37 interact with collagen via long-range electrostatic forces, initiating low-affinity binding that transitions to closerange or hydrophobic interactions. Circular dichroism analysis showed that cCBD-LL37 exhibited enhanced structural stability compared to LL37 under varying ionic strengths and pH conditions, implying potential improvements in antimicrobial activity. Moreover, we demonstrated that the release of LL37 and cCBD-LL37 into the surrounding medium was influenced by the electrostatic environment, but cCBD could enhance the retention of peptide on collagen scaffolds. Collectively, these results provide important insights into cCBD-modified AMP-binding mechanisms and suggest that the addition of cCBD may enhance peptide structural stability and retention under varying electrostatic conditions.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Characterization of LL37 Binding to Collagen through Peptide Modification with a Collagen-Binding Domain

Wei,

Rolle,

Camesano

2023

ACS Omega

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“…Moreover, in our previous in silico study, 3D structures of different KGF fusions containing CBDs from various sources were predicted, and their affinity binding to collagen and KGFR were estimated. The results showed that CBDs with medium sizes, such as CBD from Vibrio mimicus , are most appropriate for fusion to KGF, as large CBDs interfere with receptor binding, and small CBDs result in weak collagen binding 52 . Accordingly, we decided to use the CBD domain of Vibrio mimicus metalloprotease to make vibrio CBD‐KGF.…”

Section: Discussionmentioning

confidence: 99%

“…The results showed that CBDs with medium sizes, such as CBD from Vibrio mimicus , are most appropriate for fusion to KGF, as large CBDs interfere with receptor binding, and small CBDs result in weak collagen binding. 52 Accordingly, we decided to use the CBD domain of Vibrio mimicus metalloprotease to make vibrio CBD‐KGF. The CBD of Vibrio , which comprises 33 amino acids, including two essential FAXWXXT motifs, 25 was placed at the N‐terminal of KGF to increase the collagen‐binding affinity of the molecule.…”

Section: Discussionmentioning

confidence: 99%

Enhancement of keratinocyte growth factor potential in inducing adipose‐derived stem cells differentiation into keratinocytes by collagen‐targeting

Amidzadeh

Yasami-Khiabani

Rahimi

et al. 2022

J Cellular Molecular Medi

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Different growth factors can regulate stem cell differentiation. We used keratinocyte growth factor (KGF) to direct adipose-derived stem cells (ASCs) differentiation into keratinocytes. To enhance KGF bioavailability, we targeted KGF for collagen by fusing it to collagen-binding domain from Vibrio mimicus metalloprotease (vibrioCBD-KGF). KGF and vibrioCBD-KGF were expressed in Escherichia coli and purified to homogeneity. Both proteins displayed comparable activities in stimulating proliferation of HEK-293 and MCF-7 cells. vibrioCBD-KGF demonstrated enhanced collagen-binding affinity in immunofluorescence and ELISA. KGF and vibrioCBD-KGF at different concentrations (2, 10, and 20 ng/ml) were applied for 21 days on ASCs cultured on collagen-coated plates. Keratinocyte differentiation was assessed based on morphological changes, the expression of keratinocyte markers (Keratin-10 and Involucrin), and stem cell markers (Collagen-I and Vimentin) by real-time PCR or immunofluorescence. Our results indicated that the expression of keratinocyte markers was substantially increased at all concentrations of vibrioCBD-KGF, while it was observed for KGF only at 20 ng/ml. Immunofluorescence staining approved this finding. Moreover, down-regulation of Collagen-I, an indicator of differentiation commitment, was more significant in samples treated with vibrioCBD-KGF. The present study showed that vibrioCBD-KGF is more potent in inducing the ASCs differentiation into keratinocytes compared to KGF. Our results have important implications for effective skin regeneration using collagen-based biomaterials.

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Understanding the ideal wound healing mechanistic behavior using in silico modelling perspectives: A review

Abu Bakar,

Mydin,

Yusop

et al. 2024

Journal of Tissue Viability

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In silico design of fusion keratinocyte growth factor containing collagen-binding domain for tissue engineering application

Cited by 4 publications

References 67 publications

Characterization of LL37 Binding to Collagen through Peptide Modification with a Collagen-Binding Domain

Characterization of LL37 Binding to Collagen through Peptide Modification with a Collagen-Binding Domain

Enhancement of keratinocyte growth factor potential in inducing adipose‐derived stem cells differentiation into keratinocytes by collagen‐targeting

Understanding the ideal wound healing mechanistic behavior using in silico modelling perspectives: A review

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