Identification of the Kinase-Substrate Recognition Interface between MYPT1 and Rho-Kinase
Protein kinases exert physiological functions through phosphorylating their specific substrates; however, the mode of kinase–substrate recognition is not fully understood. Rho-kinase is a Ser/Thr protein kinase that regulates cytoskeletal reorganization through phosphorylating myosin light chain (ML...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-01-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/12/2/159 |
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| author | Mutsuki Amano Yoko Kanazawa Kei Kozawa Kozo Kaibuchi |
| author_facet | Mutsuki Amano Yoko Kanazawa Kei Kozawa Kozo Kaibuchi |
| author_sort | Mutsuki Amano |
| collection | DOAJ |
| description | Protein kinases exert physiological functions through phosphorylating their specific substrates; however, the mode of kinase–substrate recognition is not fully understood. Rho-kinase is a Ser/Thr protein kinase that regulates cytoskeletal reorganization through phosphorylating myosin light chain (MLC) and the myosin phosphatase targeting subunit 1 (MYPT1) of MLC phosphatase (MLCP) and is involved in various diseases, due to its aberrant cellular contraction, morphology, and movement. Despite the importance of the prediction and identification of substrates and phosphorylation sites, understanding of the precise regularity in phosphorylation preference of Rho-kinase remains far from satisfactory. Here we analyzed the Rho-kinase–MYPT1 interaction, to understand the mode of Rho-kinase substrate recognition and found that the three short regions of MYPT1 close to phosphorylation sites (referred to as docking motifs (DMs); DM1 (DLQEAEKTIGRS), DM2 (KSQPKSIRERRRPR), and DM3 (RKARSRQAR)) are important for interactions with Rho-kinase. The phosphorylation levels of MYPT1 without DMs were reduced, and the effects were limited to the neighboring phosphorylation sites. We further demonstrated that the combination of pseudosubstrate (PS) and DM of MYPT1 (PS1 + DM3 and PS2 + DM2) serves as a potent inhibitor of Rho-kinase. The present information will be useful in identifying new substrates and developing selective Rho-kinase inhibitors. |
| first_indexed | 2024-03-09T22:30:38Z |
| format | Article |
| id | doaj.art-20dcf02c1ccd48f49523a608fdd03043 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-09T22:30:38Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-20dcf02c1ccd48f49523a608fdd030432023-11-23T18:57:01ZengMDPI AGBiomolecules2218-273X2022-01-0112215910.3390/biom12020159Identification of the Kinase-Substrate Recognition Interface between MYPT1 and Rho-KinaseMutsuki Amano0Yoko Kanazawa1Kei Kozawa2Kozo Kaibuchi3Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, 65 Tsurumai, Showa-ku, Nagoya 466-8550, Aichi, JapanInstitute for Comprehensive Medical Science, Fujita Health University, Toyoake 470-1192, Aichi, JapanDepartment of Cell Pharmacology, Graduate School of Medicine, Nagoya University, 65 Tsurumai, Showa-ku, Nagoya 466-8550, Aichi, JapanDepartment of Cell Pharmacology, Graduate School of Medicine, Nagoya University, 65 Tsurumai, Showa-ku, Nagoya 466-8550, Aichi, JapanProtein kinases exert physiological functions through phosphorylating their specific substrates; however, the mode of kinase–substrate recognition is not fully understood. Rho-kinase is a Ser/Thr protein kinase that regulates cytoskeletal reorganization through phosphorylating myosin light chain (MLC) and the myosin phosphatase targeting subunit 1 (MYPT1) of MLC phosphatase (MLCP) and is involved in various diseases, due to its aberrant cellular contraction, morphology, and movement. Despite the importance of the prediction and identification of substrates and phosphorylation sites, understanding of the precise regularity in phosphorylation preference of Rho-kinase remains far from satisfactory. Here we analyzed the Rho-kinase–MYPT1 interaction, to understand the mode of Rho-kinase substrate recognition and found that the three short regions of MYPT1 close to phosphorylation sites (referred to as docking motifs (DMs); DM1 (DLQEAEKTIGRS), DM2 (KSQPKSIRERRRPR), and DM3 (RKARSRQAR)) are important for interactions with Rho-kinase. The phosphorylation levels of MYPT1 without DMs were reduced, and the effects were limited to the neighboring phosphorylation sites. We further demonstrated that the combination of pseudosubstrate (PS) and DM of MYPT1 (PS1 + DM3 and PS2 + DM2) serves as a potent inhibitor of Rho-kinase. The present information will be useful in identifying new substrates and developing selective Rho-kinase inhibitors.https://www.mdpi.com/2218-273X/12/2/159Rho-kinaseMYPT1phosphorylationsubstrate recognitioninterfacedocking motif |
| spellingShingle | Mutsuki Amano Yoko Kanazawa Kei Kozawa Kozo Kaibuchi Identification of the Kinase-Substrate Recognition Interface between MYPT1 and Rho-Kinase Biomolecules Rho-kinase MYPT1 phosphorylation substrate recognition interface docking motif |
| title | Identification of the Kinase-Substrate Recognition Interface between MYPT1 and Rho-Kinase |
| title_full | Identification of the Kinase-Substrate Recognition Interface between MYPT1 and Rho-Kinase |
| title_fullStr | Identification of the Kinase-Substrate Recognition Interface between MYPT1 and Rho-Kinase |
| title_full_unstemmed | Identification of the Kinase-Substrate Recognition Interface between MYPT1 and Rho-Kinase |
| title_short | Identification of the Kinase-Substrate Recognition Interface between MYPT1 and Rho-Kinase |
| title_sort | identification of the kinase substrate recognition interface between mypt1 and rho kinase |
| topic | Rho-kinase MYPT1 phosphorylation substrate recognition interface docking motif |
| url | https://www.mdpi.com/2218-273X/12/2/159 |
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