Crystal Structure and Substrate Specificity of PTPN12

Li, Hui; Yang, Fan; Liu, Chunhua; Peng, Xiao; Xu, Yaping; Liang, Zong-Lai; Liu, Chuan; Wang, Hongmei; Wang, Wenjun; Zheng, Wenshuai; Zhang, Wei; Ma, Xiaoyun; He, Dongfang; Song, Xiaoyuan; Cui, Fuai; Xu, Zhigang; Yi, Fan; Sun, Junying; Yu, Xiao

doi:10.1016/j.celrep.2016.04.016

Cited by 33 publications

(83 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The small-molecule inhibitors, IPA-3 and dinaciclib, were purchased from Selleck Chemicals (Houston, TX, USA). The other reagents and antibodies were the same as previously described (19).…”

Section: Antibodies and Reagentsmentioning

confidence: 99%

“…PAK1-T423E, HER2-Y1196F, GST-PTPN12-C231S, and GST-PTPN12-C231S/ S19E were generated with the Stratagene QuickChange kit (Agilent Technologies, Santa Clara, CA, USA). The other plasmids were exactly the same as previously used (19).…”

Section: Constructsmentioning

confidence: 99%

“…The methods of preparing GST-tagged protein were similar to the published description (19). Plasmids expressing GST-PTPN12 catalytic domain-transformed Escherichia coli were cultivated at 37°C until optical density reached 1.0-1.2, then, induced by 100 mM isopropyl p-D-1-thiogalactopyranoside for 16 h at 16°C.…”

Section: Protein Expression and Purificationmentioning

confidence: 99%

“…GST pull-down experiments were performed as described elsewhere (19). MDA-MB-231 cells were stimulated by 25 ng/ml epidermal growth factor (EGF) and lysed on ice after overnight starvation.…”

Section: Gst Pull-downmentioning

confidence: 99%

“…Most regulatory mechanisms of PTPN12 act through its C-terminal PEST domain, such as through the formation of protein-protein interactions via its poly-proline region or by the phosphorylation of S435 (18) or S571 (17). In addition to the previously known phosphorylation site of S39 in the catalytic domain of PTPN12 (18), we recently identified a new S19 site of PTPN12 that is phosphorylated by CDK2 (19). Previous research has demonstrated that S39 phosphorylation decreases PTPN12's enzymatic activity (18).…”

mentioning

confidence: 92%

See 4 more Smart Citations

Switching of the substrate specificity of protein tyrosine phosphatase N12 by cyclin‐dependent kinase 2 phosphorylation orchestrating 2 oncogenic pathways

Ning

et al. 2017

FASEB j.

Self Cite

View full text Add to dashboard Cite

The protein tyrosine phosphatase nonreceptor type 12 (PTPN12) is a multifunctional protein and has elicited much research attention because its decreased protein level has been associated with poor prognosis of several types of cancers. Recently, we have solved the crystal structure of the phosphatase domain of PTPN12, which disclosed a specific PTPN12-insert-loop harboring a cyclin-dependent kinase 2 (CDK2) phosphorylation site. However, the functional significance of this phosphorylation is undefined. In the present study, we found that S19 site phosphorylation of PTPN12 by CDK2 discharged its antitumor activity by down-regulation of its inhibitory role in cell migration, but not affecting its other regulatory functions. Phosphorylation of PTPN12 at the S19 site changed its substrate interface, and by doing so, selectively decreased its activity toward the human epidermal growth factor receptor 2 (HER2)- pY site, but not other HER2 phosphorylation sites or other known PTPN12 substrates. A further in-depth mechanism study revealed that the phosphorylation of PTPN12 by CDK2 impaired recruitment of the serine/threonine-protein kinase 1 (PAK1) to HER2, resulted in the blockade of the HER2-pY-PAK1-T signaling pathway, thus increased tumor cell motility. Taken together, our results identified a new phosphorylation-based substrate recognition mechanism of PTPN12 by CDK2, which orchestrated signaling crosstalk between the oncogenic CDK2 and HER2 pathways. The newly identified governing mechanism of the substrate selectivity of a particular phosphatase was previously unappreciated and exemplifies how a phospho-network is precisely controlled in different cellular contexts.-Li, H., Yang, D., Ning, S., Xu, Y., Yang, F., Yin, R., Feng, T., Han, S., Guo, L., Zhang, P., Qu, W., Guo, R., Song, C., Xiao, P., Zhou, C., Xu, Z., Sun, J.-P., Yu, X. Switching of the substrate specificity of protein tyrosine phosphatase N12 by cyclin-dependent kinase 2 phosphorylation orchestrating 2 oncogenic pathways.

show abstract

Section: Antibodies and Reagentsmentioning

confidence: 99%

Section: Constructsmentioning

confidence: 99%

Section: Protein Expression and Purificationmentioning

confidence: 99%

Section: Gst Pull-downmentioning

confidence: 99%

mentioning

confidence: 92%

See 3 more Smart Citations

Switching of the substrate specificity of protein tyrosine phosphatase N12 by cyclin‐dependent kinase 2 phosphorylation orchestrating 2 oncogenic pathways

Ning

et al. 2017

FASEB j.

Self Cite

View full text Add to dashboard Cite

show abstract

Molecular dynamics simulation studies on the specific regulation of PTPN18 to the HER2 phospho‐peptides

Tong

Liu

Sun

et al. 2021

J of Molecular Recognition

View full text Add to dashboard Cite

The specific regulation of PTPN18 protein to three HER2 phospho‐peptides has been studied by molecular dynamics simulations and free energy calculations. The results revealed that the three HER2 phospho‐peptides binding to the PTPN18 catalytic domain is energetically favorable due to substrate specificity of PTPN18, and moreover, the PTPN18 protein have significantly higher affinity to pY1248 peptide (−45.22 kcal/mol) than that of pY1112 (−25.3 kcal/mol) and pY1196 (−31.86 kcal/mol) peptides. Further, the binding of HER2 phospho‐peptides to PTPN18 have also caused the closure of WPD‐loop with the decrease of the centroid distances between the P‐loop and the WPD loop. The WPD‐loop closure of PTPN18 relates directly to the new hydrogen bond and hydrophobic interaction formations between the residues Tyr62, Asp64, Val65, Ala231, Arg235, and Ala273 in PTPN18 and Tyr(PO3) in the HER2 phospho‐peptides, which suggests that these key residues would contribute to the specific regulation of PTPN18 to the substrates. The correlation analysis revealed the allosteric communication networks from the pY binding loop to the WPD loop through the structural change and the residue interactions in PTPN18. These results will be helpful to understand the specific regulation through the allosteric communication network in the PTPN18 catalytic domain.

show abstract

In‐silico identification of Tyr232 in AMPKα2 as a dephosphorylation site for the protein tyrosine phosphatase PTP‐PEST

et al. 2023

View full text Add to dashboard Cite

The AMP‐activated protein kinase (AMPK) is known to be activated by the protein tyrosine phosphatase non‐receptor type 12 (PTP‐PEST) under hypoxic conditions. This activation is mediated by tyrosine dephosphorylation of the AMPKα subunit. However, the identity of the phosphotyrosine residues that PTP‐PEST dephosphorylates remains unknown. In this study, we first predicted the structure of the complex of the AMPKα2 subunit and PTP‐PEST catalytic domain using bioinformatics tools and further confirmed the stability of the complex using molecular dynamics simulations. Evaluation of the protein–protein interfaces indicated that residue Tyr232 is the most likely dephosphorylation site on AMPKα2. In addition, we explored the effect of phosphorylation of PTP‐PEST residue Tyr64 on the stability of the complex. Phosphorylation of the highly conserved Tyr64, an interface residue, enhances the stability of the complex via the rearrangement of a network of electrostatic interactions in conjunction with conformational changes in the catalytic WPD loop. We generated a phosphomimetic (PTP‐PEST‐Y64D) mutant and used co‐immunoprecipitation to study the effect of PTP‐PEST phosphorylation on AMPKα2 binding. The mutant exhibited an increased affinity for AMPKα2 and corroborated the in‐silico predictions. Together, our findings present a plausible structural basis of AMPK regulation by PTP‐PEST and show how phosphorylation of PTP‐PEST affects its interaction with AMPKα2.

show abstract

Crystal Structure and Substrate Specificity of PTPN12

Cited by 33 publications

References 41 publications

Switching of the substrate specificity of protein tyrosine phosphatase N12 by cyclin‐dependent kinase 2 phosphorylation orchestrating 2 oncogenic pathways

Switching of the substrate specificity of protein tyrosine phosphatase N12 by cyclin‐dependent kinase 2 phosphorylation orchestrating 2 oncogenic pathways

Molecular dynamics simulation studies on the specific regulation of PTPN18 to the HER2 phospho‐peptides

In‐silico identification of Tyr232 in AMPKα2 as a dephosphorylation site for the protein tyrosine phosphatase PTP‐PEST

Contact Info

Product

Resources

About