Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions
Summary: FHA domains are well established as phospho-dependent binding modules mediating signal transduction in Ser/Thr kinase signaling networks in both eukaryotic and prokaryotic species. Although they are unique in binding exclusively to phosphothreonine, the basis for this discrimination over p...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Elsevier B.V.
2015
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| Online Access: | http://hdl.handle.net/1721.1/96137 https://orcid.org/0000-0002-9547-3251 |
| _version_ | 1811082744415911936 |
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| author | Pennell, Simon Westcott, Sarah Ortiz-Lombardía, Miguel Patel, Dony Li, Jiejin Nott, Timothy J. Mohammed, Duaa Buxton, Roger S. Verma, Chandra Smerdon, Stephen J. Yaffe, Michael B |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Pennell, Simon Westcott, Sarah Ortiz-Lombardía, Miguel Patel, Dony Li, Jiejin Nott, Timothy J. Mohammed, Duaa Buxton, Roger S. Verma, Chandra Smerdon, Stephen J. Yaffe, Michael B |
| author_sort | Pennell, Simon |
| collection | MIT |
| description | Summary:
FHA domains are well established as phospho-dependent binding modules mediating signal transduction in Ser/Thr kinase signaling networks in both eukaryotic and prokaryotic species. Although they are unique in binding exclusively to phosphothreonine, the basis for this discrimination over phosphoserine has remained elusive. Here, we attempt to dissect overall binding specificity at the molecular level. We first determined the optimal peptide sequence for Rv0020c FHA domain binding by oriented peptide library screening. This served as a basis for systematic mutagenic and binding analyses, allowing us to derive relative thermodynamic contributions of conserved protein and peptide residues to binding and specificity. Structures of phosphopeptide-bound and uncomplexed Rv0020c FHA domain then directed molecular dynamics simulations which show how the extraordinary discrimination in favor of phosphothreonine occurs through formation of additional hydrogen-bonding networks that are ultimately stabilized by van der Waals interactions of the phosphothreonine γ-methyl group with a conserved pocket on the FHA domain surface. |
| first_indexed | 2024-09-23T12:08:19Z |
| format | Article |
| id | mit-1721.1/96137 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T12:08:19Z |
| publishDate | 2015 |
| publisher | Elsevier B.V. |
| record_format | dspace |
| spelling | mit-1721.1/961372022-10-01T08:24:24Z Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions Pennell, Simon Westcott, Sarah Ortiz-Lombardía, Miguel Patel, Dony Li, Jiejin Nott, Timothy J. Mohammed, Duaa Buxton, Roger S. Verma, Chandra Smerdon, Stephen J. Yaffe, Michael B Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Mohammed, Duaa Yaffe, Michael B. Summary: FHA domains are well established as phospho-dependent binding modules mediating signal transduction in Ser/Thr kinase signaling networks in both eukaryotic and prokaryotic species. Although they are unique in binding exclusively to phosphothreonine, the basis for this discrimination over phosphoserine has remained elusive. Here, we attempt to dissect overall binding specificity at the molecular level. We first determined the optimal peptide sequence for Rv0020c FHA domain binding by oriented peptide library screening. This served as a basis for systematic mutagenic and binding analyses, allowing us to derive relative thermodynamic contributions of conserved protein and peptide residues to binding and specificity. Structures of phosphopeptide-bound and uncomplexed Rv0020c FHA domain then directed molecular dynamics simulations which show how the extraordinary discrimination in favor of phosphothreonine occurs through formation of additional hydrogen-bonding networks that are ultimately stabilized by van der Waals interactions of the phosphothreonine γ-methyl group with a conserved pocket on the FHA domain surface. National Institutes of Health (U.S.) (GM60594) Medical Research Council (Great Britain) National Institutes of Health (U.S.) (GM68762) 2015-03-20T18:16:37Z 2015-03-20T18:16:37Z 2010-12 2010-09 Article http://purl.org/eprint/type/JournalArticle 09692126 1878-4186 http://hdl.handle.net/1721.1/96137 Pennell, Simon, Sarah Westcott, Miguel Ortiz-Lombardía, Dony Patel, Jiejin Li, Timothy J. Nott, Duaa Mohammed, et al. “Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions.” Structure 18, no. 12 (December 2010): 1587–1595. © 2010 Elsevier Ltd. https://orcid.org/0000-0002-9547-3251 en_US http://dx.doi.org/10.1016/j.str.2010.09.014 Structure Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Elsevier B.V. Elsevier |
| spellingShingle | Pennell, Simon Westcott, Sarah Ortiz-Lombardía, Miguel Patel, Dony Li, Jiejin Nott, Timothy J. Mohammed, Duaa Buxton, Roger S. Verma, Chandra Smerdon, Stephen J. Yaffe, Michael B Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions |
| title | Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions |
| title_full | Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions |
| title_fullStr | Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions |
| title_full_unstemmed | Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions |
| title_short | Structural and Functional Analysis of Phosphothreonine-Dependent FHA Domain Interactions |
| title_sort | structural and functional analysis of phosphothreonine dependent fha domain interactions |
| url | http://hdl.handle.net/1721.1/96137 https://orcid.org/0000-0002-9547-3251 |
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