In Vitro Selection of Collagen-Binding Vascular Endothelial Growth Factor Containing Genetically Encoded Mussel-Inspired Adhesive Amino Acids.
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| Formato: | Artículo científico |
| Lenguaje: | en |
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Chemistry (Weinheim an der Bergstrasse, Germany)
2025
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| author | Miwa, Takuya Yumoto, Akiko Tada, Seiichi Kim, Shin-Woong Minagawa, Noriko Matsuda, Takayoshi Ohtake, Kazumasa Shimizu, Yoshihiro Sakamoto, Kensaku Aigaki, Toshiro Ito, Yoshihiro Uzawa, Takanori |
| author_facet | Miwa, Takuya Yumoto, Akiko Tada, Seiichi Kim, Shin-Woong Minagawa, Noriko Matsuda, Takayoshi Ohtake, Kazumasa Shimizu, Yoshihiro Sakamoto, Kensaku Aigaki, Toshiro Ito, Yoshihiro Uzawa, Takanori Miwa, Takuya Yumoto, Akiko Tada, Seiichi Kim, Shin-Woong Minagawa, Noriko Matsuda, Takayoshi Ohtake, Kazumasa Shimizu, Yoshihiro Sakamoto, Kensaku Aigaki, Toshiro Ito, Yoshihiro Uzawa, Takanori |
| collection | PubMed - marine biology |
| contents | In Vitro Selection of Collagen-Binding Vascular Endothelial Growth Factor Containing Genetically Encoded Mussel-Inspired Adhesive Amino Acids. Miwa, Takuya Yumoto, Akiko Tada, Seiichi Kim, Shin-Woong Minagawa, Noriko Matsuda, Takayoshi Ohtake, Kazumasa Shimizu, Yoshihiro Sakamoto, Kensaku Aigaki, Toshiro Ito, Yoshihiro Uzawa, Takanori Collagen Vascular Endothelial Growth Factor A Animals Amino Acids Bivalvia Humans Protein Binding Dihydroxyphenylalanine Adhesives Amino Acid Sequence Peptides Protein immobilization technology is important in medical and industrial applications. We previously reported all-in-one in vitro selection, wherein a collagen-binding vascular endothelial growth factor (CB-VEGF) was identified from a fusion library of random and VEGF sequences. However, its interaction chemistry is mainly limited to the interaction established by the 20 canonical amino acids. Herein, we incorporated an adhesive non-natural amino acid found in marine mussels, L-3,4-dihydroxyphenylalanine (DOPA), into the library for all-in-one in vitro selection. After selection, we identified DOPA-containing CB-VEGF. CB-VEGF binds to collagen with an apparent dissociation constant of 2 nM; naïve VEGF does not bind to collagen. The collagen-binding peptide domain of CB-VEGF (CB-peptide) exhibited stronger binding to collagen than a mutant peptide (substitution from DOPA to tyrosine), indicating the importance of DOPA to collagen binding. The collagen-binding affinity of CB-VEGF is 10-fold higher than that of CB-peptide, suggesting that the collagen-binding ability of CB-VEGF is not due to the additive function of CB-peptide to VEGF, but is synergistic. Furthermore, increased cell growth was observed on CB-VEGF-treated collagen surfaces, not VEGF-treated collagen surfaces. Thus, integrating all-in-one in vitro selection and DOPA incorporation shows promise in generating adhesive proteins on solid supports. |
| format | Artículo científico |
| id | pubmed_39789869 |
| institution | PubMed |
| language | en |
| publishDate | 2025 |
| publisher | Chemistry (Weinheim an der Bergstrasse, Germany) |
| record_format | pubmed |
| spellingShingle | In Vitro Selection of Collagen-Binding Vascular Endothelial Growth Factor Containing Genetically Encoded Mussel-Inspired Adhesive Amino Acids. Miwa, Takuya Yumoto, Akiko Tada, Seiichi Kim, Shin-Woong Minagawa, Noriko Matsuda, Takayoshi Ohtake, Kazumasa Shimizu, Yoshihiro Sakamoto, Kensaku Aigaki, Toshiro Ito, Yoshihiro Uzawa, Takanori Collagen Vascular Endothelial Growth Factor A Animals Amino Acids Bivalvia Humans Protein Binding Dihydroxyphenylalanine Adhesives Amino Acid Sequence Peptides In Vitro Selection of Collagen-Binding Vascular Endothelial Growth Factor Containing Genetically Encoded Mussel-Inspired Adhesive Amino Acids. Miwa, Takuya Yumoto, Akiko Tada, Seiichi Kim, Shin-Woong Minagawa, Noriko Matsuda, Takayoshi Ohtake, Kazumasa Shimizu, Yoshihiro Sakamoto, Kensaku Aigaki, Toshiro Ito, Yoshihiro Uzawa, Takanori Collagen Vascular Endothelial Growth Factor A Animals Amino Acids Bivalvia Humans Protein Binding Dihydroxyphenylalanine Adhesives Amino Acid Sequence Peptides Protein immobilization technology is important in medical and industrial applications. We previously reported all-in-one in vitro selection, wherein a collagen-binding vascular endothelial growth factor (CB-VEGF) was identified from a fusion library of random and VEGF sequences. However, its interaction chemistry is mainly limited to the interaction established by the 20 canonical amino acids. Herein, we incorporated an adhesive non-natural amino acid found in marine mussels, L-3,4-dihydroxyphenylalanine (DOPA), into the library for all-in-one in vitro selection. After selection, we identified DOPA-containing CB-VEGF. CB-VEGF binds to collagen with an apparent dissociation constant of 2 nM; naïve VEGF does not bind to collagen. The collagen-binding peptide domain of CB-VEGF (CB-peptide) exhibited stronger binding to collagen than a mutant peptide (substitution from DOPA to tyrosine), indicating the importance of DOPA to collagen binding. The collagen-binding affinity of CB-VEGF is 10-fold higher than that of CB-peptide, suggesting that the collagen-binding ability of CB-VEGF is not due to the additive function of CB-peptide to VEGF, but is synergistic. Furthermore, increased cell growth was observed on CB-VEGF-treated collagen surfaces, not VEGF-treated collagen surfaces. Thus, integrating all-in-one in vitro selection and DOPA incorporation shows promise in generating adhesive proteins on solid supports. |
| title | In Vitro Selection of Collagen-Binding Vascular Endothelial Growth Factor Containing Genetically Encoded Mussel-Inspired Adhesive Amino Acids. |
| topic | Collagen Vascular Endothelial Growth Factor A Animals Amino Acids Bivalvia Humans Protein Binding Dihydroxyphenylalanine Adhesives Amino Acid Sequence Peptides |
| url | https://pubmed.ncbi.nlm.nih.gov/39789869/ |