Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2
The Tec-family kinase Btk contains a lipid-binding Pleckstrin homology and Tec homology (PH-TH) module connected by a proline-rich linker to a 'Src module', an SH3-SH2-kinase unit also found in Src-family kinases and Abl. We showed previously that Btk is activated by PH-TH dimerization, wh...
| Main Authors: | , , , , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2023
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/169560 |
| _version_ | 1826122096487432192 |
|---|---|
| author | Nocka, Laura M. Eisen, Timothy J. Iavarone, Anthony T. Groves, Jay T. Kuriyan, John |
| author2 | Institute for Digital Molecular Analytics and Science (IDMxS) |
| author_facet | Institute for Digital Molecular Analytics and Science (IDMxS) Nocka, Laura M. Eisen, Timothy J. Iavarone, Anthony T. Groves, Jay T. Kuriyan, John |
| author_sort | Nocka, Laura M. |
| collection | NTU |
| description | The Tec-family kinase Btk contains a lipid-binding Pleckstrin homology and Tec homology (PH-TH) module connected by a proline-rich linker to a 'Src module', an SH3-SH2-kinase unit also found in Src-family kinases and Abl. We showed previously that Btk is activated by PH-TH dimerization, which is triggered on membranes by the phosphatidyl inositol phosphate PIP3, or in solution by inositol hexakisphosphate (IP6) (Wang et al., 2015, https://doi.org/10.7554/eLife.06074). We now report that the ubiquitous adaptor protein growth-factor-receptor-bound protein 2 (Grb2) binds to and substantially increases the activity of PIP3-bound Btk on membranes. Using reconstitution on supported-lipid bilayers, we find that Grb2 can be recruited to membrane-bound Btk through interaction with the proline-rich linker in Btk. This interaction requires intact Grb2, containing both SH3 domains and the SH2 domain, but does not require that the SH2 domain be able to bind phosphorylated tyrosine residues - thus Grb2 bound to Btk is free to interact with scaffold proteins via the SH2 domain. We show that the Grb2-Btk interaction recruits Btk to scaffold-mediated signaling clusters in reconstituted membranes. Our findings indicate that PIP3-mediated dimerization of Btk does not fully activate Btk, and that Btk adopts an autoinhibited state at the membrane that is released by Grb2. |
| first_indexed | 2024-10-01T05:42:55Z |
| format | Journal Article |
| id | ntu-10356/169560 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:42:55Z |
| publishDate | 2023 |
| record_format | dspace |
| spelling | ntu-10356/1695602023-07-26T15:32:03Z Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2 Nocka, Laura M. Eisen, Timothy J. Iavarone, Anthony T. Groves, Jay T. Kuriyan, John Institute for Digital Molecular Analytics and Science (IDMxS) Science::Biological sciences Protein-Tyrosine Kinases Dimerization The Tec-family kinase Btk contains a lipid-binding Pleckstrin homology and Tec homology (PH-TH) module connected by a proline-rich linker to a 'Src module', an SH3-SH2-kinase unit also found in Src-family kinases and Abl. We showed previously that Btk is activated by PH-TH dimerization, which is triggered on membranes by the phosphatidyl inositol phosphate PIP3, or in solution by inositol hexakisphosphate (IP6) (Wang et al., 2015, https://doi.org/10.7554/eLife.06074). We now report that the ubiquitous adaptor protein growth-factor-receptor-bound protein 2 (Grb2) binds to and substantially increases the activity of PIP3-bound Btk on membranes. Using reconstitution on supported-lipid bilayers, we find that Grb2 can be recruited to membrane-bound Btk through interaction with the proline-rich linker in Btk. This interaction requires intact Grb2, containing both SH3 domains and the SH2 domain, but does not require that the SH2 domain be able to bind phosphorylated tyrosine residues - thus Grb2 bound to Btk is free to interact with scaffold proteins via the SH2 domain. We show that the Grb2-Btk interaction recruits Btk to scaffold-mediated signaling clusters in reconstituted membranes. Our findings indicate that PIP3-mediated dimerization of Btk does not fully activate Btk, and that Btk adopts an autoinhibited state at the membrane that is released by Grb2. Published version This work was supported by the National Institutes of Health (grant number PO1 A1091580) 2023-07-24T07:36:29Z 2023-07-24T07:36:29Z 2023 Journal Article Nocka, L. M., Eisen, T. J., Iavarone, A. T., Groves, J. T. & Kuriyan, J. (2023). Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2. ELife, 12, e82676-. https://dx.doi.org/10.7554/eLife.82676 2050-084X https://hdl.handle.net/10356/169560 10.7554/eLife.82676 37159508 2-s2.0-85157984380 12 e82676 en PO1 A1091580 eLife © 2023 Nocka et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. application/pdf |
| spellingShingle | Science::Biological sciences Protein-Tyrosine Kinases Dimerization Nocka, Laura M. Eisen, Timothy J. Iavarone, Anthony T. Groves, Jay T. Kuriyan, John Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2 |
| title | Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2 |
| title_full | Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2 |
| title_fullStr | Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2 |
| title_full_unstemmed | Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2 |
| title_short | Stimulation of the catalytic activity of the tyrosine kinase Btk by the adaptor protein Grb2 |
| title_sort | stimulation of the catalytic activity of the tyrosine kinase btk by the adaptor protein grb2 |
| topic | Science::Biological sciences Protein-Tyrosine Kinases Dimerization |
| url | https://hdl.handle.net/10356/169560 |
| work_keys_str_mv | AT nockalauram stimulationofthecatalyticactivityofthetyrosinekinasebtkbytheadaptorproteingrb2 AT eisentimothyj stimulationofthecatalyticactivityofthetyrosinekinasebtkbytheadaptorproteingrb2 AT iavaroneanthonyt stimulationofthecatalyticactivityofthetyrosinekinasebtkbytheadaptorproteingrb2 AT grovesjayt stimulationofthecatalyticactivityofthetyrosinekinasebtkbytheadaptorproteingrb2 AT kuriyanjohn stimulationofthecatalyticactivityofthetyrosinekinasebtkbytheadaptorproteingrb2 |