A selective Seoul-Fluor-based bioprobe, SfBP, for vaccinia H1-related phosphatase—a dual-specific protein tyrosine phosphatase

Jeong, Myeong Seon; Kim, Eunha; Kang, Hyo Jin; Choi, Eun Joung; Cho, Alvin R.; Chung, Sang J.; Park, Seung Bum

doi:10.1039/c2cc31377d

Cited by 23 publications

(9 citation statements)

References 26 publications

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“…DUSP26-N exhibited significantly higher phosphatase activity than DUSP26-C at different concentrations, whereas DUSP26-N (C152S) was catalytically inactive against DiFMUP (Fig 1B). In addition, we performed enzyme kinetics analysis with DUSP26-N and compared the kinetic parameters with those of VH1-related DUSP (VHR), DUSP13b, and DUSP14 [27] (Table 1). The K cat /K m value of DUSP26-N was 100-fold smaller than that of VHR, but was comparable to that of DUSP13b, which is known as the DUSP that is most analogous to DUSP26 according to phylogenetic analysis [2, 28].…”

Section: Resultsmentioning

confidence: 99%

Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26

Won

Lee

et al. 2016

PLoS ONE

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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.

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Section: Resultsmentioning

confidence: 99%

Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26

Won

Lee

et al. 2016

PLoS ONE

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“…Furthermore, we demonstrated that the emission wavelength of Seoul‐Fluors can be predicted by computational analysis of the energy gap between the excited and ground states 4b. Because of the ability to predict the emission wavelength, we were able to develop a ratiometric pH sensor,5a a turn‐on lipid‐droplet bioprobe,5b,c and a selective turn‐off probe of the dual‐specific phosphatase DUSP3 5d. With these studies, we clearly demonstrated the importance of the substituents at the C7 and C9 positions in perturbing the photophysical properties of Seoul‐Fluors.…”

Section: Methodsmentioning

confidence: 97%

Design and Preparation of a Lithium‐rich Layered Oxide Cathode with a Mg‐Concentration‐Gradient Shell for Improved Rate Capability

Wang

et al. 2015

ChemElectroChem

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Fluorescence imaging enables the uniquely sensitive observation of functional‐ and molecular‐recognition events in living cells. However, only a limited range of biological processes have been subjected to imaging because of the lack of a design strategy and difficulties in the synthesis of biosensors. Herein, we report a facile synthesis of emission‐tunable and predictable Seoul‐Fluors, 9‐aryl‐1,2‐dihydrolopyrrolo[3,4‐b]indolizin‐3‐ones, with various R1 and R2 substituents by coinage‐metal‐catalyzed intramolecular 1,3‐dipolar cycloaddition and subsequent palladium‐mediated CH activation. We also showed that the quantum yields of Seoul‐Fluors are controlled by the electronic nature of the substituents, which influences the extent of photoinduced electron transfer. On the basis of this understanding, we demonstrated our design strategy by the development of a Seoul‐Fluor‐based chemosensor 20 for reactive oxygen species that was not accessible by a previous synthetic route.

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“…SHP2 was overexpressed in RosettaTM (DE3) and purified using a metal affinity resin ( Figure S1 in Supplementary Materials). The catalytic activity of the purified proteins was evaluated using DiFMUP as a fluorogenic substrate, which has been widely used to determine PTP activity [25]. The k cat , K M , and k cat /K M values were determined to be 270 min −1 , 70 µM, and 3.9 µM −1 min −1 , respectively, using the Lineweaver-Burk plot analysis (Table S1 in Supplementary Materials).…”

Section: Expression Purification and Kinetic Characterization Of Shp2mentioning

confidence: 99%

Polyphyllin D Shows Anticancer Effect through a Selective Inhibition of Src Homology Region 2-Containing Protein Tyrosine Phosphatase-2 (SHP2)

et al. 2021

Self Cite

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Natural products have continued to offer tremendous opportunities for drug development, as they have long been used in traditional medicinal systems. SHP2 has served as an anticancer target. To identify novel SHP2 inhibitors with potential anticancer activity, we screened a library containing 658 natural products. Polyphyllin D was found to selectively inhibit SHP2 over SHP1, whereas two other identified compounds (echinocystic acid and oleanolic acid) demonstrated dual SHP1 and SHP2 inhibition. In a cell-based assay, polyphyllin D exhibited cytotoxicity in Jurkat cells, an acute lymphoma leukemia cell line, whereas the other two compounds were ineffective. Polyphyllin D also decreased the level of phosphorylated extracellular signal-regulated kinase (p-ERK), a proliferation marker in Jurkat cells. Furthermore, knockdown of protein tyrosine phosphatase (PTP)N6 (SHP1) or PTPN11 (SHP2) decreased p-ERK levels. However, concurrent knockdown of PTPN6 and PTPN11 in Jurkat cells recovered p-ERK levels. These results demonstrated that polyphyllin D has potential anticancer activity, which can be attributed to its selective inhibition of SHP2 over SHP1.

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A selective Seoul-Fluor-based bioprobe, SfBP, for vaccinia H1-related phosphatase—a dual-specific protein tyrosine phosphatase

Cited by 23 publications

References 26 publications

Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26

Structural Insight into the Critical Role of the N-Terminal Region in the Catalytic Activity of Dual-Specificity Phosphatase 26

Design and Preparation of a Lithium‐rich Layered Oxide Cathode with a Mg‐Concentration‐Gradient Shell for Improved Rate Capability

Polyphyllin D Shows Anticancer Effect through a Selective Inhibition of Src Homology Region 2-Containing Protein Tyrosine Phosphatase-2 (SHP2)

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