Biochemical and Cellular Evidence Demonstrating AKT-1 as a Binding Partner for Resveratrol Targeting Protein NQO2

Abstract

BackgroundAKT plays an important role in the control of cell proliferation and survival. Aberrant activation of AKT frequently occurs in human cancers making it an attractive drug targets and leading to the synthesis of numerous AKT inhibitors as therapeutic candidates. Less is known regarding proteins that control AKT. We recently reported that quinone reductase 2 (NQO2) inhibited AKT activity, by unknown mechanisms.Methodology/Principal FindingsIn this study, molecular modeling was used to query interaction between NQO2 and AKT. We found that pleckstrin homology (PH) and kinase domains of AKT bind to chains A and B of NQO2. Pull-down and deletion assays revealed that PH domain of AKT is essential for interaction with NQO2. Modeling analysis further revealed that kinase domain of AKT binds NQO2 in the vicinity of asparagine 161 located in the resveratrol-binding domain of NQO2. In studies to test whether exposure to resveratrol potentiates or diminishes AKT binding to NQO2, we showed that pre-binding by resveratrol in wild type but not histidine-161 (N161H) mutant NQO2 significantly affected this interaction. To obtain information on interplay between resveratrol and AKT, resveratrol affinity chromatography was performed. AKT binds with high affinity to the column suggesting that it is a target of resveratrol. The half-life of AKT mRNA decreased from ∼4 h in control cells to ∼1 h in NQO2-knockdown cells. The inhibition of AKT by resveratrol was attenuated in NQO2-expressing relative to NQO2-knockdown cells.Conclusion/SignificanceBoth NQO2 and AKT are targets of resveratrol; NQO2:AKT interaction is a novel physiological regulator of AKT activation/function.

TXNIP regulates AKT‐mediated cellular senescence by direct interaction under glucose‐mediated metabolic stress

“…Protein kinase C interacts with the PH domain of Bruton tyrosine kinase (BTK) and inhibits BTK activity (Yao, Kawakami, & Kawakami, 1994). One group has reported that N‐ribosyldihydronicotinamide:quinone reductase 2 (NQO2), an oxidoreductive enzyme, interacts with AKT via the PH domain of AKT to inhibit AKT activity (Hsieh et al, 2014). Here, we showed that TXNIP interacts with AKT via the PH domain of AKT and inhibits the kinase activity of AKT (Figure 3).…”

“…AKT is a serine–threonine kinase that is involved in a variety of cellular processes including cell survival, proliferation, and metabolism (Hsieh, Lin, Bennett, Wu, & Wu, 2014). AKT has a structure consisting of an N‐terminal pleckstrin homology (PH) domain, a kinase domain (KD), and a C‐terminal hydrophobic regulatory region (HM) (Parikh et al, 2012).…”

NRH:quinone oxidoreductase 2 (NQO2) and glutaminase (GLS) both play a role in large extracellular vesicles (LEV) formation in preclinical LNCaP‐C4‐2B prostate cancer model of progressive metastasis

“…Apart from Asian men, incidence of prostate cancer among men is also significantly low in France which is due, a significant part, to a large intake of red wine, a natural source of resveratrol, leading to the concept of “French Paradox.” Work from our laboratories have focused for many years on the molecular mechanisms of androgen independence and bone metastasis in prostate cancer, with particular reference to the use of curcumin (used in Asian cuisine) and resveratrol (found in red wine) as significant anti‐inflammatory, anti‐oxidative agents, and as nutritional modulators of cancer progression . Our earlier work on the mechanisms of androgen independence in prostate cancer cells has led to a better understanding of the function of the enzyme NRH: quinone oxidoreductase (NQO2) in prostate carcinogenesis . In this communication, the role of the enzyme NQO2 is investigated further with respect to its role not only in regulating oxidative stress but also in regulating prostate cancer metastasis and how glutamine metabolism influences all these processes.…”

“…[20][21][22][23][24] Our earlier work on the mechanisms of androgen independence in prostate cancer cells has led to a better understanding of the function of the enzyme NRH: quinone oxidoreductase (NQO2) in prostate carcinogenesis. 25,26 In this communication, the role of the enzyme NQO2 is investigated further with respect to its role not only in regulating oxidative stress but also in regulating prostate cancer metastasis and how glutamine metabolism influences all these processes. Our results highly suggest a heretofore uninvestigated role for the enzyme NQO2 in the genesis and function of large extracellular vesicles (LEVs), particularly in the modulation of the bony microenvironment in advanced prostate cancer through its specific interaction with caveolin-1 which is implicated in the genesis of hormone refractory and metastatic prostate cancer.…”

Application of Mixture Design Response Surface Methodology for Combination Chemotherapy in PC-3 Human Prostate Cancer Cells

“…Many naturally occurring compounds such as resveratrol (RES), flavopiridol (FLAV), and piperlongumine (PIP) are multimechanistic compounds that have been identified as putative chemotherapeutics (Pan et al, 2012). Treatment with RES has been shown to cause increases in cytosolic calcium (Ma et al, 2007;Peterson et al, 2016), bind to various proteins to induce intrinsic apoptosis (Locatelli et al, 2005;Calamini et al, 2010;Hsieh et al, 2014), and cause production of ROS (Miki et al, 2012). FLAV inhibits cyclin dependent kinases to disrupt the cell cycle and causes production of ROS (Gupta et al, 2010;Shao et al, 2016), whereas PIP has been shown to cause ROS production and inhibit signal transducer and activator of transcription 3 and nuclear factor kappa-light-chain-enhancer of activated B cells (Kong et al, 2008;Adams et al, 2012;Ginzburg et al, 2014;Gong et al, 2014;Bharadwaj et al, 2015).…”