Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26.
Human dual-specificity phosphatase 26 (DUSP26) is a novel target for anticancer therapy because its dephosphorylation of the p53 tumor suppressor regulates the apoptosis of cancer cells. DUSP26 inhibition results in neuroblastoma cell cytotoxicity through p53-mediated apoptosis. Despite the previous...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2016-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC5008780?pdf=render |
| _version_ | 1828530672676896768 |
|---|---|
| author | Eun-Young Won Sang-Ok Lee Dong-Hwa Lee Daeyoup Lee Kwang-Hee Bae Sang Chul Lee Seung Jun Kim Seung-Wook Chi |
| author_facet | Eun-Young Won Sang-Ok Lee Dong-Hwa Lee Daeyoup Lee Kwang-Hee Bae Sang Chul Lee Seung Jun Kim Seung-Wook Chi |
| author_sort | Eun-Young Won |
| collection | DOAJ |
| description | Human dual-specificity phosphatase 26 (DUSP26) is a novel target for anticancer therapy because its dephosphorylation of the p53 tumor suppressor regulates the apoptosis of cancer cells. DUSP26 inhibition results in neuroblastoma cell cytotoxicity through p53-mediated apoptosis. Despite the previous structural studies of DUSP26 catalytic domain (residues 61-211, DUSP26-C), the high-resolution structure of its catalytically active form has not been resolved. In this study, we determined the crystal structure of a catalytically active form of DUSP26 (residues 39-211, DUSP26-N) with an additional N-terminal region at 2.0 Å resolution. Unlike the C-terminal domain-swapped dimeric structure of DUSP26-C, the DUSP26-N (C152S) monomer adopts a fold-back conformation of the C-terminal α8-helix and has an additional α1-helix in the N-terminal region. Consistent with the canonically active conformation of its protein tyrosine phosphate-binding loop (PTP loop) observed in the structure, the phosphatase assay results demonstrated that DUSP26-N has significantly higher catalytic activity than DUSP26-C. Furthermore, size exclusion chromatography-multiangle laser scattering (SEC-MALS) measurements showed that DUSP26-N (C152S) exists as a monomer in solution. Notably, the crystal structure of DUSP26-N (C152S) revealed that the N-terminal region of DUSP26-N (C152S) serves a scaffolding role by positioning the surrounding α7-α8 loop for interaction with the PTP-loop through formation of an extensive hydrogen bond network, which seems to be critical in making the PTP-loop conformation competent for phosphatase activity. Our study provides the first high-resolution structure of a catalytically active form of DUSP26, which will contribute to the structure-based rational design of novel DUSP26-targeting anticancer therapeutics. |
| first_indexed | 2024-12-11T22:27:28Z |
| format | Article |
| id | doaj.art-2f53f4b17f10401b9fe3d7b83acff29c |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T22:27:28Z |
| publishDate | 2016-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-2f53f4b17f10401b9fe3d7b83acff29c2022-12-22T00:48:15ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01119e016211510.1371/journal.pone.0162115Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26.Eun-Young WonSang-Ok LeeDong-Hwa LeeDaeyoup LeeKwang-Hee BaeSang Chul LeeSeung Jun KimSeung-Wook ChiHuman dual-specificity phosphatase 26 (DUSP26) is a novel target for anticancer therapy because its dephosphorylation of the p53 tumor suppressor regulates the apoptosis of cancer cells. DUSP26 inhibition results in neuroblastoma cell cytotoxicity through p53-mediated apoptosis. Despite the previous structural studies of DUSP26 catalytic domain (residues 61-211, DUSP26-C), the high-resolution structure of its catalytically active form has not been resolved. In this study, we determined the crystal structure of a catalytically active form of DUSP26 (residues 39-211, DUSP26-N) with an additional N-terminal region at 2.0 Å resolution. Unlike the C-terminal domain-swapped dimeric structure of DUSP26-C, the DUSP26-N (C152S) monomer adopts a fold-back conformation of the C-terminal α8-helix and has an additional α1-helix in the N-terminal region. Consistent with the canonically active conformation of its protein tyrosine phosphate-binding loop (PTP loop) observed in the structure, the phosphatase assay results demonstrated that DUSP26-N has significantly higher catalytic activity than DUSP26-C. Furthermore, size exclusion chromatography-multiangle laser scattering (SEC-MALS) measurements showed that DUSP26-N (C152S) exists as a monomer in solution. Notably, the crystal structure of DUSP26-N (C152S) revealed that the N-terminal region of DUSP26-N (C152S) serves a scaffolding role by positioning the surrounding α7-α8 loop for interaction with the PTP-loop through formation of an extensive hydrogen bond network, which seems to be critical in making the PTP-loop conformation competent for phosphatase activity. Our study provides the first high-resolution structure of a catalytically active form of DUSP26, which will contribute to the structure-based rational design of novel DUSP26-targeting anticancer therapeutics.http://europepmc.org/articles/PMC5008780?pdf=render |
| spellingShingle | Eun-Young Won Sang-Ok Lee Dong-Hwa Lee Daeyoup Lee Kwang-Hee Bae Sang Chul Lee Seung Jun Kim Seung-Wook Chi Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26. PLoS ONE |
| title | Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26. |
| title_full | Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26. |
| title_fullStr | Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26. |
| title_full_unstemmed | Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26. |
| title_short | Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26. |
| title_sort | structural insight into the critical role of the n terminal region in the catalytic activity of dual specificity phosphatase 26 |
| url | http://europepmc.org/articles/PMC5008780?pdf=render |
| work_keys_str_mv | AT eunyoungwon structuralinsightintothecriticalroleofthenterminalregioninthecatalyticactivityofdualspecificityphosphatase26 AT sangoklee structuralinsightintothecriticalroleofthenterminalregioninthecatalyticactivityofdualspecificityphosphatase26 AT donghwalee structuralinsightintothecriticalroleofthenterminalregioninthecatalyticactivityofdualspecificityphosphatase26 AT daeyouplee structuralinsightintothecriticalroleofthenterminalregioninthecatalyticactivityofdualspecificityphosphatase26 AT kwangheebae structuralinsightintothecriticalroleofthenterminalregioninthecatalyticactivityofdualspecificityphosphatase26 AT sangchullee structuralinsightintothecriticalroleofthenterminalregioninthecatalyticactivityofdualspecificityphosphatase26 AT seungjunkim structuralinsightintothecriticalroleofthenterminalregioninthecatalyticactivityofdualspecificityphosphatase26 AT seungwookchi structuralinsightintothecriticalroleofthenterminalregioninthecatalyticactivityofdualspecificityphosphatase26 |