Structural basis for the binding of the cancer targeting scorpion toxin, ClTx, to the vascular endothelia growth factor receptor neuropilin-1
Chlorotoxin (ClTx) is a 36-residue disulfide-rich peptide isolated from the venom of the scorpion Leiurus quinquestriatus. This peptide has been shown to selectively bind to brain tumours (gliomas), however, with conflicting reports regarding its direct cellular target. Recently, the vascular endoth...
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| Format: | Article |
| Language: | English |
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Elsevier
2021-01-01
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| Series: | Current Research in Structural Biology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2665928X21000155 |
| _version_ | 1798026729204219904 |
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| author | Gagan Sharma Carolyne B. Braga Kai-En Chen Xinying Jia Venkatraman Ramanujam Brett M. Collins Roberto Rittner Mehdi Mobli |
| author_facet | Gagan Sharma Carolyne B. Braga Kai-En Chen Xinying Jia Venkatraman Ramanujam Brett M. Collins Roberto Rittner Mehdi Mobli |
| author_sort | Gagan Sharma |
| collection | DOAJ |
| description | Chlorotoxin (ClTx) is a 36-residue disulfide-rich peptide isolated from the venom of the scorpion Leiurus quinquestriatus. This peptide has been shown to selectively bind to brain tumours (gliomas), however, with conflicting reports regarding its direct cellular target. Recently, the vascular endothelial growth factor receptor, neuropilin-1 (NRP1) has emerged as a potential target of the peptide. Here, we sought to characterize the details of the binding of ClTx to the b1-domain of NRP1 (NRP1-b1) using solution state nuclear magnetic resonance (NMR) spectroscopy. The 3D structure of the isotope labelled peptide was solved using multidimensional heteronuclear NMR spectroscopy to produce a well-resolved structural ensemble. The structure points to three putative protein-protein interaction interfaces, two basic patches (R14/K15/K23 and R25/K27/R36) and a hydrophobic patch (F6/T7/T8/H10). The NRP1-b1 binding interface of ClTx was elucidated using 15N chemical shift mapping and included the R25/K27/R36 region of the peptide. The thermodynamics of binding was determined using isothermal titration calorimetry (ITC). In both NMR and ITC measurements, the binding was shown to be competitive with a known NRP1-b1 inhibitor. Finally, combining all of this data we generate a model of the ClTx:NRP1-b1 complex. The data shows that the peptide binds to the same region of NRP1 that is used by the SARS-CoV-2 virus for cell entry, however, via a non-canonical binding mode. Our results provide evidence for a non-standard NRP1 binding motif, while also providing a basis for further engineering of ClTx to generate peptides with improved NRP1 binding for future biomedical applications. |
| first_indexed | 2024-04-11T18:40:03Z |
| format | Article |
| id | doaj.art-af52d3404ddf4994b4ff881afbc923be |
| institution | Directory Open Access Journal |
| issn | 2665-928X |
| language | English |
| last_indexed | 2024-04-11T18:40:03Z |
| publishDate | 2021-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Current Research in Structural Biology |
| spelling | doaj.art-af52d3404ddf4994b4ff881afbc923be2022-12-22T04:09:03ZengElsevierCurrent Research in Structural Biology2665-928X2021-01-013179186Structural basis for the binding of the cancer targeting scorpion toxin, ClTx, to the vascular endothelia growth factor receptor neuropilin-1Gagan Sharma0Carolyne B. Braga1Kai-En Chen2Xinying Jia3Venkatraman Ramanujam4Brett M. Collins5Roberto Rittner6Mehdi Mobli7Centre for Advanced Imaging, The University of Queensland, St Lucia, QLD, AustraliaCentre for Advanced Imaging, The University of Queensland, St Lucia, QLD, Australia; Chemistry Institute, University of Campinas, P.O. Box 6154, 13083-970, Campinas, SP, BrazilInstitute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, AustraliaCentre for Advanced Imaging, The University of Queensland, St Lucia, QLD, AustraliaCentre for Advanced Imaging, The University of Queensland, St Lucia, QLD, AustraliaInstitute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, AustraliaChemistry Institute, University of Campinas, P.O. Box 6154, 13083-970, Campinas, SP, BrazilCentre for Advanced Imaging, The University of Queensland, St Lucia, QLD, Australia; Corresponding author.Chlorotoxin (ClTx) is a 36-residue disulfide-rich peptide isolated from the venom of the scorpion Leiurus quinquestriatus. This peptide has been shown to selectively bind to brain tumours (gliomas), however, with conflicting reports regarding its direct cellular target. Recently, the vascular endothelial growth factor receptor, neuropilin-1 (NRP1) has emerged as a potential target of the peptide. Here, we sought to characterize the details of the binding of ClTx to the b1-domain of NRP1 (NRP1-b1) using solution state nuclear magnetic resonance (NMR) spectroscopy. The 3D structure of the isotope labelled peptide was solved using multidimensional heteronuclear NMR spectroscopy to produce a well-resolved structural ensemble. The structure points to three putative protein-protein interaction interfaces, two basic patches (R14/K15/K23 and R25/K27/R36) and a hydrophobic patch (F6/T7/T8/H10). The NRP1-b1 binding interface of ClTx was elucidated using 15N chemical shift mapping and included the R25/K27/R36 region of the peptide. The thermodynamics of binding was determined using isothermal titration calorimetry (ITC). In both NMR and ITC measurements, the binding was shown to be competitive with a known NRP1-b1 inhibitor. Finally, combining all of this data we generate a model of the ClTx:NRP1-b1 complex. The data shows that the peptide binds to the same region of NRP1 that is used by the SARS-CoV-2 virus for cell entry, however, via a non-canonical binding mode. Our results provide evidence for a non-standard NRP1 binding motif, while also providing a basis for further engineering of ClTx to generate peptides with improved NRP1 binding for future biomedical applications.http://www.sciencedirect.com/science/article/pii/S2665928X21000155ChlorotoxinNRP1Disulfide-rich peptideHeteronuclear NMRITC |
| spellingShingle | Gagan Sharma Carolyne B. Braga Kai-En Chen Xinying Jia Venkatraman Ramanujam Brett M. Collins Roberto Rittner Mehdi Mobli Structural basis for the binding of the cancer targeting scorpion toxin, ClTx, to the vascular endothelia growth factor receptor neuropilin-1 Current Research in Structural Biology Chlorotoxin NRP1 Disulfide-rich peptide Heteronuclear NMR ITC |
| title | Structural basis for the binding of the cancer targeting scorpion toxin, ClTx, to the vascular endothelia growth factor receptor neuropilin-1 |
| title_full | Structural basis for the binding of the cancer targeting scorpion toxin, ClTx, to the vascular endothelia growth factor receptor neuropilin-1 |
| title_fullStr | Structural basis for the binding of the cancer targeting scorpion toxin, ClTx, to the vascular endothelia growth factor receptor neuropilin-1 |
| title_full_unstemmed | Structural basis for the binding of the cancer targeting scorpion toxin, ClTx, to the vascular endothelia growth factor receptor neuropilin-1 |
| title_short | Structural basis for the binding of the cancer targeting scorpion toxin, ClTx, to the vascular endothelia growth factor receptor neuropilin-1 |
| title_sort | structural basis for the binding of the cancer targeting scorpion toxin cltx to the vascular endothelia growth factor receptor neuropilin 1 |
| topic | Chlorotoxin NRP1 Disulfide-rich peptide Heteronuclear NMR ITC |
| url | http://www.sciencedirect.com/science/article/pii/S2665928X21000155 |
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