Structural and functional characterization of the human protein kinase ASK1.
Apoptosis signal-regulating kinase 1 (ASK1) plays an essential role in stress and immune response and has been linked to the development of several diseases. Here, we present the structure of the human ASK1 catalytic domain in complex with staurosporine. Analytical ultracentrifugation (AUC) and crys...
| Main Authors: | , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2007
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| Subjects: |
| _version_ | 1797061469964075008 |
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| author | Bunkoczi, G Salah, E Filippakopoulos, P Fedorov, O Müller, S Sobott, F Parker, S Zhang, H Min, W Turk, B Knapp, S |
| author_facet | Bunkoczi, G Salah, E Filippakopoulos, P Fedorov, O Müller, S Sobott, F Parker, S Zhang, H Min, W Turk, B Knapp, S |
| author_sort | Bunkoczi, G |
| collection | OXFORD |
| description | Apoptosis signal-regulating kinase 1 (ASK1) plays an essential role in stress and immune response and has been linked to the development of several diseases. Here, we present the structure of the human ASK1 catalytic domain in complex with staurosporine. Analytical ultracentrifugation (AUC) and crystallographic analysis showed that ASK1 forms a tight dimer (K(d) approximately 0.2 microM) interacting in a head-to-tail fashion. We found that the ASK1 phosphorylation motifs differ from known ASK1 phosphorylation sites but correspond well to autophosphorylation sites identified by mass spectrometry. Reporter gene assays showed that all three identified in vitro autophosphorylation sites (Thr813, Thr838, Thr842) regulate ASK1 signaling, but site-directed mutants showed catalytic activities similar to wild-type ASK1, suggesting a regulatory mechanism independent of ASK1 kinase activity. The determined high-resolution structure of ASK1 and identified ATP mimetic inhibitors will provide a first starting point for the further development of selective inhibitors. |
| first_indexed | 2024-03-06T20:31:37Z |
| format | Journal article |
| id | oxford-uuid:313b870e-a272-4371-9731-e81092434631 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T20:31:37Z |
| publishDate | 2007 |
| record_format | dspace |
| spelling | oxford-uuid:313b870e-a272-4371-9731-e810924346312022-03-26T13:06:40ZStructural and functional characterization of the human protein kinase ASK1.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:313b870e-a272-4371-9731-e81092434631Enzyme InhibitorsAmino Acid SequencechemistryHumansStaurosporineModels, MolecularMolecular Sequence DataPhosphorylationBinding SitesProtein ConformationMAP Kinase Kinase Kinase 5CatalysismetabolismProtein Structure, TertiaryDimerizationSubstrate SpecificityEnzyme ActivationStructure-Activity RelationshipEnglishStructural Genomics Consortium2007Bunkoczi, GSalah, EFilippakopoulos, PFedorov, OMüller, SSobott, FParker, SZhang, HMin, WTurk, BKnapp, SApoptosis signal-regulating kinase 1 (ASK1) plays an essential role in stress and immune response and has been linked to the development of several diseases. Here, we present the structure of the human ASK1 catalytic domain in complex with staurosporine. Analytical ultracentrifugation (AUC) and crystallographic analysis showed that ASK1 forms a tight dimer (K(d) approximately 0.2 microM) interacting in a head-to-tail fashion. We found that the ASK1 phosphorylation motifs differ from known ASK1 phosphorylation sites but correspond well to autophosphorylation sites identified by mass spectrometry. Reporter gene assays showed that all three identified in vitro autophosphorylation sites (Thr813, Thr838, Thr842) regulate ASK1 signaling, but site-directed mutants showed catalytic activities similar to wild-type ASK1, suggesting a regulatory mechanism independent of ASK1 kinase activity. The determined high-resolution structure of ASK1 and identified ATP mimetic inhibitors will provide a first starting point for the further development of selective inhibitors. |
| spellingShingle | Enzyme Inhibitors Amino Acid Sequence chemistry Humans Staurosporine Models, Molecular Molecular Sequence Data Phosphorylation Binding Sites Protein Conformation MAP Kinase Kinase Kinase 5 Catalysis metabolism Protein Structure, Tertiary Dimerization Substrate Specificity Enzyme Activation Structure-Activity Relationship Bunkoczi, G Salah, E Filippakopoulos, P Fedorov, O Müller, S Sobott, F Parker, S Zhang, H Min, W Turk, B Knapp, S Structural and functional characterization of the human protein kinase ASK1. |
| title | Structural and functional characterization of the human protein kinase ASK1. |
| title_full | Structural and functional characterization of the human protein kinase ASK1. |
| title_fullStr | Structural and functional characterization of the human protein kinase ASK1. |
| title_full_unstemmed | Structural and functional characterization of the human protein kinase ASK1. |
| title_short | Structural and functional characterization of the human protein kinase ASK1. |
| title_sort | structural and functional characterization of the human protein kinase ask1 |
| topic | Enzyme Inhibitors Amino Acid Sequence chemistry Humans Staurosporine Models, Molecular Molecular Sequence Data Phosphorylation Binding Sites Protein Conformation MAP Kinase Kinase Kinase 5 Catalysis metabolism Protein Structure, Tertiary Dimerization Substrate Specificity Enzyme Activation Structure-Activity Relationship |
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