Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells
Three artificial proteins that bind the gadolinium ion (Gd<sup>3+</sup>) with tumour-specific ligands were de novo engineered and tested as candidate drugs for binary radiotherapy (BRT) and contrast agents for magnetic resonance imaging (MRI). Gd<sup>3+</sup>-binding modules...
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MDPI AG
2022-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/6/3297 |
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| author | Natalia V. Pozdniakova Oxana V. Ryabaya Alevtina S. Semkina Vsevolod A. Skribitsky Alexei B. Shevelev |
| author_facet | Natalia V. Pozdniakova Oxana V. Ryabaya Alevtina S. Semkina Vsevolod A. Skribitsky Alexei B. Shevelev |
| author_sort | Natalia V. Pozdniakova |
| collection | DOAJ |
| description | Three artificial proteins that bind the gadolinium ion (Gd<sup>3+</sup>) with tumour-specific ligands were de novo engineered and tested as candidate drugs for binary radiotherapy (BRT) and contrast agents for magnetic resonance imaging (MRI). Gd<sup>3+</sup>-binding modules were derived from calmodulin. They were joined with elastin-like polypeptide (ELP) repeats from human elastin to form the four-centre Gd<sup>3+</sup>-binding domain (4MBS-domain) that further was combined with F3 peptide (a ligand of nucleolin, a tumour marker) to form the F3-W4 block. The F3-W4 block was taken alone (E2-13W4 protein), as two repeats (E1-W8) and as three repeats (E1-W12). Each protein was supplemented with three copies of the RGD motif (a ligand of integrin αvβ3) and green fluorescent protein (GFP). In contrast to Magnevist (a Gd-containing contrast agent), the proteins exhibited three to four times higher accumulation in U87MG glioma and A375 melanoma cell lines than in normal fibroblasts. The proteins remained for >24 h in tumours induced by Ca755 adenocarcinoma in C57BL/6 mice. They exhibited stability towards blood proteases and only accumulated in the liver and kidney. The technological advantages of using the engineered proteins as a basis for developing efficient and non-toxic agents for early diagnosis of tumours by MRI as well as part of BRT were demonstrated. |
| first_indexed | 2024-03-09T19:41:09Z |
| format | Article |
| id | doaj.art-2ba66034ecdd499bae20cdc96fe893e5 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T19:41:09Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-2ba66034ecdd499bae20cdc96fe893e52023-11-24T01:36:57ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-03-01236329710.3390/ijms23063297Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour CellsNatalia V. Pozdniakova0Oxana V. Ryabaya1Alevtina S. Semkina2Vsevolod A. Skribitsky3Alexei B. Shevelev4N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Public Health of the Russian Federation (N.N. Blokhin NMRCO), Kashirskoe Shosse, 23, 115478 Moscow, RussiaN.N. Blokhin National Medical Research Center of Oncology of the Ministry of Public Health of the Russian Federation (N.N. Blokhin NMRCO), Kashirskoe Shosse, 23, 115478 Moscow, RussiaDepartment of Medical Nanobiotechnologies, Pirogov Russian National Research Medical University, Ostrovityanova, 1, 117997 Moscow, RussiaN.N. Blokhin National Medical Research Center of Oncology of the Ministry of Public Health of the Russian Federation (N.N. Blokhin NMRCO), Kashirskoe Shosse, 23, 115478 Moscow, RussiaN.I. Vavilov Institute of General Genetics RAS, Gubkina 3, GSP-1, 119991 Moscow, RussiaThree artificial proteins that bind the gadolinium ion (Gd<sup>3+</sup>) with tumour-specific ligands were de novo engineered and tested as candidate drugs for binary radiotherapy (BRT) and contrast agents for magnetic resonance imaging (MRI). Gd<sup>3+</sup>-binding modules were derived from calmodulin. They were joined with elastin-like polypeptide (ELP) repeats from human elastin to form the four-centre Gd<sup>3+</sup>-binding domain (4MBS-domain) that further was combined with F3 peptide (a ligand of nucleolin, a tumour marker) to form the F3-W4 block. The F3-W4 block was taken alone (E2-13W4 protein), as two repeats (E1-W8) and as three repeats (E1-W12). Each protein was supplemented with three copies of the RGD motif (a ligand of integrin αvβ3) and green fluorescent protein (GFP). In contrast to Magnevist (a Gd-containing contrast agent), the proteins exhibited three to four times higher accumulation in U87MG glioma and A375 melanoma cell lines than in normal fibroblasts. The proteins remained for >24 h in tumours induced by Ca755 adenocarcinoma in C57BL/6 mice. They exhibited stability towards blood proteases and only accumulated in the liver and kidney. The technological advantages of using the engineered proteins as a basis for developing efficient and non-toxic agents for early diagnosis of tumours by MRI as well as part of BRT were demonstrated.https://www.mdpi.com/1422-0067/23/6/3297recombinant protein engineeringELPbinary radiotherapygadolinium contrast agentF3 peptideRGD |
| spellingShingle | Natalia V. Pozdniakova Oxana V. Ryabaya Alevtina S. Semkina Vsevolod A. Skribitsky Alexei B. Shevelev Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells International Journal of Molecular Sciences recombinant protein engineering ELP binary radiotherapy gadolinium contrast agent F3 peptide RGD |
| title | Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells |
| title_full | Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells |
| title_fullStr | Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells |
| title_full_unstemmed | Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells |
| title_short | Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells |
| title_sort | using elp repeats as a scaffold for de novo construction of gadolinium binding domains within multifunctional recombinant proteins for targeted delivery of gadolinium to tumour cells |
| topic | recombinant protein engineering ELP binary radiotherapy gadolinium contrast agent F3 peptide RGD |
| url | https://www.mdpi.com/1422-0067/23/6/3297 |
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