Structural Insights into the Active Site Formation of DUSP22 in N-loop-containing Protein Tyrosine Phosphatases
| Autor: | Tse-Hua Tan, Ping-Chiang Lyu, Huai-Chia Chuang, Chih-Hsuan Lai, Co-Chih Chang |
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| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular DUSP22 Protein Conformation Stereochemistry Phosphatase DUSPs Protein tyrosine phosphatase Cys-based PTPs Article Catalysis lcsh:Chemistry Inorganic Chemistry Serine Dephosphorylation N-loop Catalytic Domain Humans Protein Interaction Domains and Motifs Amino Acid Sequence Enzyme kinetics Amino Acids Physical and Theoretical Chemistry lcsh:QH301-705.5 Molecular Biology Conserved Sequence Spectroscopy Substrate Interaction Binding Sites biology Chemistry Organic Chemistry Active site Hydrogen Bonding General Medicine active site hydrogen bonding network Computer Science Applications lcsh:Biology (General) lcsh:QD1-999 Mutation biology.protein Dual-Specificity Phosphatases Mitogen-Activated Protein Kinase Phosphatases Protein Tyrosine Phosphatases Protein Binding Cysteine |
| Zdroj: | International Journal of Molecular Sciences Volume 21 Issue 20 International Journal of Molecular Sciences, Vol 21, Iss 7515, p 7515 (2020) |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms21207515 |
| Popis: | Cysteine-based protein tyrosine phosphatases (Cys-based PTPs) perform dephosphorylation to regulate signaling pathways in cellular responses. The hydrogen bonding network in their active site plays an important conformational role and supports the phosphatase activity. Nearly half of dual-specificity phosphatases (DUSPs) use three conserved residues, including aspartate in the D-loop, serine in the P-loop, and asparagine in the N-loop, to form the hydrogen bonding network, the D-, P-, N-triloop interaction (DPN&ndash triloop interaction). In this study, DUSP22 is used to investigate the importance of the DPN&ndash triloop interaction in active site formation. Alanine mutations and somatic mutations of the conserved residues, D57, S93, and N128 substantially decrease catalytic efficiency (kcat/KM) by more than 102-fold. Structural studies by NMR and crystallography reveal that each residue can perturb the three loops and induce conformational changes, indicating that the hydrogen bonding network aligns the residues in the correct positions for substrate interaction and catalysis. Studying the DPN&ndash triloop interaction reveals the mechanism maintaining phosphatase activity in N-loop-containing PTPs and provides a foundation for further investigation of active site formation in different members of this protein class. |
| Databáze: | OpenAIRE |
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