How the biotin-streptavidin interaction was made even stronger: investigation via crystallography and a chimaeric tetramer.
The interaction between SA (streptavidin) and biotin is one of the strongest non-covalent interactions in Nature. SA is a widely used tool and a paradigm for protein-ligand interactions. We previously developed a SA mutant, termed Tr (traptavidin), possessing a 10-fold lower off-rate for biotin, wit...
| Main Authors: | , , , |
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| Format: | Journal article |
| Language: | English |
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2011
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| _version_ | 1797061456482533376 |
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| author | Chivers, C Koner, A Lowe, E Howarth, M |
| author_facet | Chivers, C Koner, A Lowe, E Howarth, M |
| author_sort | Chivers, C |
| collection | OXFORD |
| description | The interaction between SA (streptavidin) and biotin is one of the strongest non-covalent interactions in Nature. SA is a widely used tool and a paradigm for protein-ligand interactions. We previously developed a SA mutant, termed Tr (traptavidin), possessing a 10-fold lower off-rate for biotin, with increased mechanical and thermal stability. In the present study, we determined the crystal structures of apo-Tr and biotin-Tr at 1.5 Å resolution. In apo-SA the loop (L3/4), near biotin's valeryl tail, is typically disordered and open, but closes upon biotin binding. In contrast, L3/4 was shut in both apo-Tr and biotin-Tr. The reduced flexibility of L3/4 and decreased conformational change on biotin binding provide an explanation for Tr's reduced biotin off- and on-rates. L3/4 includes Ser45, which forms a hydrogen bond to biotin consistently in Tr, but erratically in SA. Reduced breakage of the biotin-Ser45 hydrogen bond in Tr is likely to inhibit the initiating event in biotin's dissociation pathway. We generated a Tr with a single biotin-binding site rather than four, which showed a simi-larly low off-rate, demonstrating that Tr's low off-rate was governed by intrasubunit effects. Understanding the structural features of this tenacious interaction may assist the design of even stronger affinity tags and inhibitors. |
| first_indexed | 2024-03-06T20:31:25Z |
| format | Journal article |
| id | oxford-uuid:312b2cbb-a782-40c9-9cea-1e43ffa8a4cc |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T20:31:25Z |
| publishDate | 2011 |
| record_format | dspace |
| spelling | oxford-uuid:312b2cbb-a782-40c9-9cea-1e43ffa8a4cc2022-03-26T13:06:11ZHow the biotin-streptavidin interaction was made even stronger: investigation via crystallography and a chimaeric tetramer.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:312b2cbb-a782-40c9-9cea-1e43ffa8a4ccEnglishSymplectic Elements at Oxford2011Chivers, CKoner, ALowe, EHowarth, MThe interaction between SA (streptavidin) and biotin is one of the strongest non-covalent interactions in Nature. SA is a widely used tool and a paradigm for protein-ligand interactions. We previously developed a SA mutant, termed Tr (traptavidin), possessing a 10-fold lower off-rate for biotin, with increased mechanical and thermal stability. In the present study, we determined the crystal structures of apo-Tr and biotin-Tr at 1.5 Å resolution. In apo-SA the loop (L3/4), near biotin's valeryl tail, is typically disordered and open, but closes upon biotin binding. In contrast, L3/4 was shut in both apo-Tr and biotin-Tr. The reduced flexibility of L3/4 and decreased conformational change on biotin binding provide an explanation for Tr's reduced biotin off- and on-rates. L3/4 includes Ser45, which forms a hydrogen bond to biotin consistently in Tr, but erratically in SA. Reduced breakage of the biotin-Ser45 hydrogen bond in Tr is likely to inhibit the initiating event in biotin's dissociation pathway. We generated a Tr with a single biotin-binding site rather than four, which showed a simi-larly low off-rate, demonstrating that Tr's low off-rate was governed by intrasubunit effects. Understanding the structural features of this tenacious interaction may assist the design of even stronger affinity tags and inhibitors. |
| spellingShingle | Chivers, C Koner, A Lowe, E Howarth, M How the biotin-streptavidin interaction was made even stronger: investigation via crystallography and a chimaeric tetramer. |
| title | How the biotin-streptavidin interaction was made even stronger: investigation via crystallography and a chimaeric tetramer. |
| title_full | How the biotin-streptavidin interaction was made even stronger: investigation via crystallography and a chimaeric tetramer. |
| title_fullStr | How the biotin-streptavidin interaction was made even stronger: investigation via crystallography and a chimaeric tetramer. |
| title_full_unstemmed | How the biotin-streptavidin interaction was made even stronger: investigation via crystallography and a chimaeric tetramer. |
| title_short | How the biotin-streptavidin interaction was made even stronger: investigation via crystallography and a chimaeric tetramer. |
| title_sort | how the biotin streptavidin interaction was made even stronger investigation via crystallography and a chimaeric tetramer |
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