Broad-spectrum chemokine-binding peptides from class A evasins
The chemokine network comprises 46 secreted chemokines that signal through 18 GPCRs, resulting in leukocyte recruitment in inflammation. Despite being a validated therapeutic target, chemokine network redundancy thwarts pharmacological intervention. Class A and Class B tick salivary proteins (evasin...
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| Other Authors: | |
| Format: | Thesis |
| Language: | English |
| Published: |
2024
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| _version_ | 1826317558259646464 |
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| author | Vales, S |
| author2 | Bhattacharya, S |
| author_facet | Bhattacharya, S Vales, S |
| author_sort | Vales, S |
| collection | OXFORD |
| description | The chemokine network comprises 46 secreted chemokines that signal through 18 GPCRs, resulting in leukocyte recruitment in inflammation. Despite being a validated therapeutic target, chemokine network redundancy thwarts pharmacological intervention. Class A and Class B tick salivary proteins (evasins) bind CC and CXC-class chemokines, respectively, overcoming redundancy and allowing prolonged blood feeding. Evasins are unsuitable for therapeutic use due to potential immunogenicity and chemokine-class specificity requiring both evasin classes to be used. Peptides previously isolated from the chemokine-binding region of class A evasins inhibit a limited number of CC-class chemokines but lack the breadth of binding needed to overcome redundancy. This thesis tests the hypothesises that phage display can identify and improve peptides from class A evasins with broad chemokine binding and inhibitory activity. By screening a phage-display library from 21 class A evasins, peptides that surprisingly bind both CC and CXC chemokine classes were identified. An exemplar peptide, HD2, from evasin EVA4, inhibited both CC and CXC- class chemokines. Alanine-scanning mutagenesis of HD2 identified critical residues important for inhibitory activity, which correlated with binding activity in phage display. HD845 is a peptide derived from the evasin EV672. HD845 binds and inhibits both CC and CXC-chemokines in phage display and in chemotaxis assays. Saturation mutagenesis of HD845 identified several single mutations that show enhanced chemokine binding and inhibitory activity. Lastly, HD2 served as a template to identify improved mutant peptides capable of inhibiting an atherosclerosis-relevant chemokine pool. These results support the hypothesis that peptides with broad-spectrum chemokine-binding activity can be identified, and their binding affinity and potency improved using phage display. |
| first_indexed | 2025-03-11T16:55:48Z |
| format | Thesis |
| id | oxford-uuid:d1ba1003-4a7d-407e-a853-11a715d480ef |
| institution | University of Oxford |
| language | English |
| last_indexed | 2025-03-11T16:55:48Z |
| publishDate | 2024 |
| record_format | dspace |
| spelling | oxford-uuid:d1ba1003-4a7d-407e-a853-11a715d480ef2025-02-20T15:07:39ZBroad-spectrum chemokine-binding peptides from class A evasinsThesishttp://purl.org/coar/resource_type/c_db06uuid:d1ba1003-4a7d-407e-a853-11a715d480efInflammationChemokineEnglishHyrax Deposit2024Vales, SBhattacharya, SRedwood, CDavies, GThe chemokine network comprises 46 secreted chemokines that signal through 18 GPCRs, resulting in leukocyte recruitment in inflammation. Despite being a validated therapeutic target, chemokine network redundancy thwarts pharmacological intervention. Class A and Class B tick salivary proteins (evasins) bind CC and CXC-class chemokines, respectively, overcoming redundancy and allowing prolonged blood feeding. Evasins are unsuitable for therapeutic use due to potential immunogenicity and chemokine-class specificity requiring both evasin classes to be used. Peptides previously isolated from the chemokine-binding region of class A evasins inhibit a limited number of CC-class chemokines but lack the breadth of binding needed to overcome redundancy. This thesis tests the hypothesises that phage display can identify and improve peptides from class A evasins with broad chemokine binding and inhibitory activity. By screening a phage-display library from 21 class A evasins, peptides that surprisingly bind both CC and CXC chemokine classes were identified. An exemplar peptide, HD2, from evasin EVA4, inhibited both CC and CXC- class chemokines. Alanine-scanning mutagenesis of HD2 identified critical residues important for inhibitory activity, which correlated with binding activity in phage display. HD845 is a peptide derived from the evasin EV672. HD845 binds and inhibits both CC and CXC-chemokines in phage display and in chemotaxis assays. Saturation mutagenesis of HD845 identified several single mutations that show enhanced chemokine binding and inhibitory activity. Lastly, HD2 served as a template to identify improved mutant peptides capable of inhibiting an atherosclerosis-relevant chemokine pool. These results support the hypothesis that peptides with broad-spectrum chemokine-binding activity can be identified, and their binding affinity and potency improved using phage display. |
| spellingShingle | Inflammation Chemokine Vales, S Broad-spectrum chemokine-binding peptides from class A evasins |
| title | Broad-spectrum chemokine-binding peptides from class A evasins |
| title_full | Broad-spectrum chemokine-binding peptides from class A evasins |
| title_fullStr | Broad-spectrum chemokine-binding peptides from class A evasins |
| title_full_unstemmed | Broad-spectrum chemokine-binding peptides from class A evasins |
| title_short | Broad-spectrum chemokine-binding peptides from class A evasins |
| title_sort | broad spectrum chemokine binding peptides from class a evasins |
| topic | Inflammation Chemokine |
| work_keys_str_mv | AT valess broadspectrumchemokinebindingpeptidesfromclassaevasins |