Small molecule activator of Nm23/NDPK as an inhibitor of metastasis

Lee, Jae Jin; Kim, Hwang Suk; Lee, Ji Sun; Park, Jimin; Shin, Sang Chul; Song, Soonwha; Lee, Eun-Sun; Choi, Jung Eun; Suh, Ji Wan; Lee, Hongsoo; Kim, Eunice Eun Kyeong; Seo, Eun Kyoung; Shin, Dongkun; Lee, Ho‐Young; Lee, Hee Yoon; Lee, Kong Joo

doi:10.1038/s41598-018-29101-6

Cited by 18 publications

(15 citation statements)

References 41 publications

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“…The biochemical assays and several cell biology assays were also conducted using NME2 (Nm23-H2) with similar results. NME2 has been reported to have both similarities and differences with NME1 (48,49), but in this case the data are generalizable. NME1 and NME2 are also known to form heterooligomers that could influence their interactome, subcellular localization, and possibly their function.…”

Section: Discussionmentioning

confidence: 64%

Metastasis Suppressors NME1 and NME2 Promote Dynamin 2 Oligomerization and Regulate Tumor Cell Endocytosis, Motility, and Metastasis

2019

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NM23 (NME) is a metastasis suppressor that significantly reduces metastasis without affecting primary tumor size, however, the precise molecular mechanisms are not completely understood. We examined the role of dynamin (DNM2), a GTPase regulating membrane scission of vesicles in endocytosis, in NME1 and NME2 regulation of tumor cell motility and metastasis. Overexpression of NMEs in MDA-MB-231T and MDA-MB-435 cancer cell lines increased endocytosis of transferrin and EGF receptors (TfR and EGFR) concurrent with motility and migration suppression. The internalized vesicles, costained with Rab5, had AP2 depleted from the cell surface and exhibited increased Rab5-GTP levels, consistent with endocytosis. Dynamin inhibitors Iminodyn-22 and Dynole-34-2, or shRNA-mediated downregulation of DNM2, impaired NME's ability to augment endocytosis or suppress tumor cell motility. In a lung metastasis assay, NME1 overexpression failed to significantly suppress metastasis in the DNM2 knockdown MDA-MB-231T cells. Using the EGF-EGFR signaling axis as a model in MDA-MB-231T cells, NME1 decreased pEGFR and pAkt expression in a DNM2-dependent manner, indicating the relevance of this interaction for downstream signaling. NME-DNM2 interaction was confirmed in two-way coimmunoprecipitations. Transfection of a NME1 site-directed mutant lacking histidine protein kinase activity but retaining nucleoside diphosphate kinase (NDPK) activity showed that the NDPK activity of NME was insufficient to promote endocytosis or inhibit EGFR signaling. We show that addition of NME1 or NME2 to DNM2 facilitates DNM2 oligomerization and increases GTPase activity, both required for vesicle scission. NME-DNM2 interaction may contribute to metastasis suppression by altering tumor endocytic and motility phenotypes. Significance: NME1 suppresses metastasis via changes in tumor endocytosis and motility, mediated by dynamin (DNM2) GTPase activity.

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Section: Discussionmentioning

confidence: 64%

Metastasis Suppressors NME1 and NME2 Promote Dynamin 2 Oligomerization and Regulate Tumor Cell Endocytosis, Motility, and Metastasis

2019

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“…There have been many efforts to characterize protein structures that have the key to elucidating the complex protein function and dynamics. Recently, protein structural changes by oxidation of peroxiredoxin 2 23 , by calcium binding in EF-hands of secretagogin 24 , and by chemical binding to Nm23-H1 25 and by drug binding to PPARγ 26 have all been well characterized by employing HDX-MS. HDX-MS is a one of the hottest techniques, which costs less effort and money, and at the same time, it has less limitation compared to other methods such as NMR or X-ray crystallography as described previously 27 . With the growing data size and the technology, a fully automated method is essential for high throughput analysis.…”

Section: Discussionmentioning

confidence: 99%

deMix: Decoding Deuterated Distributions from Heterogeneous Protein States via HDX-MS

Lee

Joo

et al. 2019

Sci Rep

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Characterization of protein structural changes in response to protein modifications, ligand or chemical binding, or protein-protein interactions is essential for understanding protein function and its regulation. Amide hydrogen/deuterium exchange (HDX) coupled with mass spectrometry (MS) is one of the most favorable tools for characterizing the protein dynamics and changes of protein conformation. However, currently the analysis of HDX-MS data is not up to its full power as it still requires manual validation by mass spectrometry experts. Especially, with the advent of high throughput technologies, the data size grows everyday and an automated tool is essential for the analysis. Here, we introduce a fully automated software, referred to as ‘deMix’, for the HDX-MS data analysis. deMix deals directly with the deuterated isotopic distributions, but not considering their centroid masses and is designed to be robust over random noises. In addition, unlike the existing approaches that can only determine a single state from an isotopic distribution, deMix can also detect a bimodal deuterated distribution, arising from EX1 behavior or heterogeneous peptides in conformational isomer proteins. Furthermore, deMix comes with visualization software to facilitate validation and representation of the analysis results.

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“…It binds to the C-terminus of Nm23-H1 to stabilize the hexamer structure and induces NDPK activation. To identify the binding region of NMac1 with Nm23-H1 and its mode of action, discernible changes were identified in three peptic peptides, aa 2–8, 64–75, and 142–152 residues, in a time-dependent manner by using HDX-MS 101 . Among these three peptides, hydrogen-deuterium exchanges in two peptides (aa 2–8 and 142–152) were significantly decreased by NMac1 binding, and these regions formed a small pocket in the C-terminal of Nm23-H1, indicating that NMac1 interacts with the C-terminus of Nm23-H1.…”

Section: Nm23-h1 As a Metastasis Suppressormentioning

confidence: 99%

Activation of Nm23-H1 to suppress breast cancer metastasis via redox regulation

Lee

2021

Exp Mol Med

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Non-metastatic protein 23 H1 (Nm23-H1), a housekeeping enzyme, is a nucleoside diphosphate kinase-A (NDPK-A). It was the first identified metastasis suppressor protein. Nm23-H1 prolongs disease-free survival and is associated with a good prognosis in breast cancer patients. However, the molecular mechanisms underlying the role of Nm23-H1 in biological processes are still not well understood. This is a review of recent studies focusing on controlling NDPK activity based on the redox regulation of Nm23-H1, structural, and functional changes associated with the oxidation of cysteine residues, and the relationship between NDPK activity and cancer metastasis. Further understanding of the redox regulation of the NDPK function will likely provide a new perspective for developing new strategies for the activation of NDPK-A in suppressing cancer metastasis.

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Small molecule activator of Nm23/NDPK as an inhibitor of metastasis

Cited by 18 publications

References 41 publications

Metastasis Suppressors NME1 and NME2 Promote Dynamin 2 Oligomerization and Regulate Tumor Cell Endocytosis, Motility, and Metastasis

Metastasis Suppressors NME1 and NME2 Promote Dynamin 2 Oligomerization and Regulate Tumor Cell Endocytosis, Motility, and Metastasis

deMix: Decoding Deuterated Distributions from Heterogeneous Protein States via HDX-MS

Activation of Nm23-H1 to suppress breast cancer metastasis via redox regulation

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