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Targeting adenine nucleotide translocase-2 (ANT2) to overcome resistance to epidermal growth factor receptor tyrosine kinase inhibitor in non-small cell lung cancer Ji Young Jang, Yong Goo Kim, Soo Jeong Nam, Bhumsuk Keam , Tae Min Kim , Yoon Kyung Jeon , Chul Woo Kim 41. NFV treatment inhibited . ANT1 is involved not only in the processes of ADP/ATP exchange but also in the composition of the mitochondrial membrane permeability transition pore (mPTP); and the function of ANT1 is closely related to its own conformational . From: Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2017 Download as PDF About this page Programmed Cardiomyocyte Death in Heart Disease Adenine nucleotide translocase (ANT) is an integral protein located in the inner mitochondrial membrane and plays an important role in maintaining ATP/ADP ratio, thereby contributing to the energy-supplying function of mitochondria. Molecular Formula: C 31 H 46 O 18 S 2 •xK. Adenine nucleotide translocase-2 (ANT2), one of the four adenine nucleotide translocase isoforms expressed in humans, is expressed at high levels in undifferentiated cells and tissues with high pro-liferating and regenerating capacity, including lympho-cytes, kidney, and liver [14-16]. The first family, which includes atractyloside (ATR) and carboxyatractyloside (CATR), binds to the ADP/ATP translocase from the cytoplasmic side, locking it in a cytoplasmic side open conformation. The net accumulation of adenine nucleotides was inhibited 76 ± 1.8% by 100 μm N-ethylmaleimide and to a lesser extent (57 ± 0.5%) by 50 μm atractyloside. Similar to the knockdown of adenine nucleotide translocase-2, genistein treatment decreased ADP uptake into the mitochondria and ATP synthesis. Adenine nucleotide translocase-2 (ANT2) is an oncogenic mitochondrial membrane-associated protein. AU - Aprille, June R. Light-activated verteporfin-induced apoptosis was abolished by transfection with Bcl-2, a procedure reported to inhibit the mitochondrial permeability transition pore complex (PTPC). membrane proteins, translocator protein (TSPO) and adenine nucleotide translocase 2 (ANT2), are both considered to be components of PTP complex and are overexpressed in cancers compared to normal tissues. However, patients eventually develop resistance to EGFR-TKIs by several mechanisms. the adenine nucleotide-binding site of the translocase and that this binding is facilitated, and the local con- centration of the inhibitor increased, by the binding of the hydrocarbon chain of the fatty acid moiety (non-polar end of acyl-CoA molecule) to mitochon- drial membrane. GSAO (4-(N-(S-glutathionylacetyl)amino) phenylarsonous acid) and PENAO (4-(N-(S-penicillaminylacetyl)amino) phenylarsonous acid) are tumour metabolism inhibitors that target adenine nucleotide translocase (ANT) of the inner-mitochondrial membrane. Two alternative mechanisms are studied: 1), dynamic compartmentation of ATP and ADP, . The net accumulation of adenine nucleotides was inhibited 76 ± 1.8% by 100 μm N-ethylmaleimide and to a lesser extent (57 ± 0.5%) by 50 μm atractyloside. Furthermore, by applying flux control theory to these inhibitor titrations, it was possible to demonstrate that the adenine nucleotide translocase exerted greater control over respiration in the newborn than in the adult, and at maximal rates of coupled respiration the translocase had a control strength of 0.98. Biosynthesis of the mitochondrial adenine nucleotide translocase (ATPase) inhibitor bongkrekic acid in Burkholderia gladioli† Barbara Rohm , a Kirstin Scherlach a and Christian Hertweck * ab 15, 1387-1396. doi: 10.1158/1535-7163.mct-15-0089 Bongkrekic acid is a specific inhibitor of the adenine nucleotide translocase (ANT) and blocks the MPT in isolated mitochondria or cells (39, 41, 42). A P Halestrap, A M Davidson; Inhibition of Ca 2+-induced large-amplitude swelling of liver and heart mitochondria by cyclosporin is probably caused by the inhibitor binding to mitochondrial-matrix peptidyl-prolyl cis-trans isomerase and preventing it interacting with the adenine nucleotide translocase. Concentrations of 1 to 25 pM inhibitor reduce the ATP hydrolysis rate about 25%. Using mitochondria from rat liver, which lack uncoupling proteins, in the present study we . Crossref, Medline, Google Scholar; Zamora M, Merono C, Vinas O, Mampel T (2004b). We report that the photosensitizer verteporfin kills lymphoma cells by an apoptotic process involving a dissipation of the mitochondrial inner transmembrane potential (ΔΨm). Hwang et al. In addition, ANT can cooperate with Bax to form a lethal pore during apoptosis. Adenine nucleotide translocase is an ADP/ATP transporter in the inner mitochondrial membrane. A Novel Adenine Nucleotide Translocase Inhibitor, MT-21, Induces Cytochrome c Release by a Mitochondrial Permeability Transition-independent Mechanism* The release of cytochrome c from mitochondria is a critical step during apoptosis. Adenine nucleotide translocator (ANT) 2 is highly expressed in proliferative cells, and ANT2 induction in cancer cells is known to be directly associated with glycolytic metabolisms and carcinogenesis. View this article via: PubMed CrossRef Google Scholar. Adenine nucleotide translocase-1 (ANT1) is a mitochondrial inner membrane protein located in the mitochondria and is responsible for mitochondrial ADP/ATP transport (Parodi-Rullaán et al, 2019; Ruprecht & Kunji, 2020 ). Supposing complete inhibition of the adenine nucleotide translocase, one may conclude that about 75% of the ATP-ase is associated with inside-out membranes. This process, known as permeability transition, evokes severe dysfunction in mitochondria through the opening of a non-specific pore, whose chemical nature is still under discussion. Note: One important exception to this solubilization Mild uncoupling of mitochondria is known to be cardioprotective, and adenine nucleotide translocase 1 (ANT1) is a key mediator of mitochondrial uncoupling. The trivalent arsenical moiety of GSAO and PENAO reacts with two matrix . However, patients eventually develop resistance to EGFR-TKIs by several mechanisms. Application: Carboxyatractyloside potassium salt is a highly selective inhibitor of adenine nucleotide translocase (ANT) CAS Number: 35988-42-2 (non-salt) Purity: ≥98%. AU - Aprille, June R. These proteins all appear to have molecular masses in the 50-60-kDa range. ADP:ATP antiporter that mediates import of ADP into the mitochondrial matrix for ATP synthesis, and export of ATP out to fuel the cell (By similarity). Humans possess four distinct ANT isoforms, encoded by four genes . 1990. Adenine nucleotide translocator - Inhibition. Halestrap AP, Davidson AM. Mutations within Ant1 have been shown to produce a syndrome of chronic progressive external ophthalmoplegia (CPEO) in humans. ANT1 is involved not only in the processes of ADP/ATP exchange but also in the composition of the mitochondrial membrane permeability transition pore (mPTP); and the function of ANT1 is closely related to its own conformational changes. Adenine nucleotide translocase (ANT) is known as a core component of the mitochondrial permeability transition pore (MPTP) and a key player in cell death. RuR also prevented PTP opening promoted by atractyloside, an adenine nucleotide translocase inhibitor. Moreover, coimmunoprecipitation and pulldown analyses confirmed that the ORF4 protein interacts directly with mitochondrial ANT3 (mtANT3). To validate the specificity of this assay, a number of known compounds with defined mode of action were tested, which include mitochondrial complex inhibitors, membrane . Adenine nucleotide translocase-1 (ANT1) is an ADP/ATP transporter protein located in the inner mitochondrial membrane. Mersalyl (10 nmol/mg protein) did not inhibit efflux. Adenine nucleotide translocase-2 exchanges ADP/ATP through the mitochondrial inner membrane. However, its role in camptothecin (CPT)-induced apoptosis has not been examined. Adenine nucleotide translocator (ANT), also known as the ADP/ATP translocase (ANT), ADP/ATP carrier protein (AAC) or mitochondrial ADP/ATP carrier, exchanges free ATP with free ADP across the inner mitochondrial membrane. These results suggest that VDAC, located in the mitochondrial outer membrane, controls Ca(2 . Figure 1D shows RLM undergoing sOMF. ANT, adenine nucleotide translocase; TTFA, thenoyltrifluoroacetone; FCCP, carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone; MPTP, mitochondrial permeability transition pore. . Using thermodynamic assumptions and computer modeling, we established that mitochondrial membrane potential can be more negative than the reversal potential of the adenine nucleotide translocase (ANT) but more positive than that of the F 0 F1-ATPase. The protein contains a C-MYC/DDK Tag. ANT1 also contains redox-sensitive cysteines that may be targets for modification by nitroalkenes. In addition, IMAC is also able to transport AMP, while the adenine nucleotide translo- J. Biol. Adenine Nucleotide Translocase Detection. Ant also possesses a discrete membrane uncoupling activity. Adenine nucleotide translocase (Ant) is the most abundant protein on the mitochondrial inner membrane (MIM) primarily involved in ADP/ATP exchange. It has been well documented [3,5,13,14] that atractylate, bongkrekic acid and long-chain acyl CoA esters inhibit the transport of adenine nucleotides 4r -I PROTEIN (mg) Fig.1. There are some proposals regarding the components of the pore structure, e.g., the adenine nucleotide translocase and dimers of the F1 Fo-ATP synthase. which mediates K' uniport (18-20) and is inhibited by ATP (20). Inhibition of Ca2(+)-induced large-amplitude swelling of liver and heart mitochondria by cyclosporin is probably caused by the inhibitor binding to mitochondrial-matrix peptidyl-prolyl cis-trans isomerase and preventing it interacting with the adenine nucleotide translocase. Adenine Nucleotide Translocator ANT plays a central role in cell bioenergetics by exporting ATP from mitochondrial matrix to cytosol and in regulating the ADP/ATP ratio in mitochondrial OXPHOS. Adenine nucleotide translocator ( ANT ), also known as the ADP/ATP translocase ( ANT ), ADP/ATP carrier protein ( AAC) or mitochondrial ADP/ATP carrier, exchanges free ATP with free ADP across the inner mitochondrial membrane. Biochem J. Agaric acid is used to regulate lipid metabolism. 1290 Yet another model, initially proposed by our group, predicts that the adenine nucleotide translocase (ANT) in the inner membrane can form a channel which, upon opening, triggers matrix swelling . ADP/ATP translocase is very specifically inhibited by two families of compounds. It also mediates mitochondria-mediated cell death through interactions with Bax and Bcl-2, pro- and anti . Recently, ANT2 was Supplemental Information: This is classified as a Dangerous Good for transport and may be subject to . Agaric acid promotes efflux of accumulated Ca 2+, collapse of transmembrane potential, and mitochondrial swelling. Mol. Adenine nucleotide translocase (ANT) is a nuclear-encoded protein abundantly located in the inner mitochondrial mem-brane, and the role of this protein is to catalyze the exchange of mitochondrial ATP with cytosolic ADP. ANT-1 is a component of the mitochondrial permeability transition complex, a protein aggregate connecting the inner with the outer mitochondrial membrane that has recently been implicated in apoptosis. Subsequently, ORF4 protein colocalization with adenine nucleotide translocase 3 (ANT3) was observed using structured illumination microscopy. Chem. Mitochondria generate reactive oxygen species, whose downstream lipid peroxidation products, such as 4-hydroxynonenal, induce uncoupling of oxidative phosphorylation by increasing proton leak through mitochondrial inner membrane proteins such as the uncoupling proteins and adenine nucleotide translocase. The adenine nucleotide translocase: a central component of the mitochondrial permeability transition pore and key player in cell death. 272:3346-3354. ApppI inhibited the mitochondrial ADP/ATP translocase and caused apoptosis in osteoclasts. Mitochondrial adenine nucleotide translocase (ANT) is believed to be a component or a regulatory component of the mitochondrial permeability transition pore (mtPTP), which controls mitochondrial permeability transition during apoptosis. The data provide further support for our hypothesis that lipid-protein interactions are important determinants in the activity of the adenine nucleotide translocase in mitochondria. Mitochondrial membrane carriers containing proline and cysteine, such as adenine nucleotide translocase (ANT), are potential targets of cyclophilin D (CyP-D) and potential Ca2+-induced permeability transition pore (PTP) components or regulators; CyP-D, a mitochondrial peptidyl-prolyl cis-trans isomerase, is the probable target of the PTP inhibitor cyclosporine A (CsA). Therefore it is the key link between ATP production in the mitochondrial matrix space and ATP consumption in the cytosol. . , including the well-known ADP/ATP translocase (1). The adenine nucleotide transporter 1 ( Ant1) gene encodes an inner mitochondrial membrane protein that transports ADP into mitochondria and ATP from mitochondria to the cytosol. These effects induced the opening of the permeability transition pore and release of cytochrome c, by its interaction with a component of the non-specific pore complex, fixed to the carrier in the conformation c, as carboxyatractyloside does. The adenine nucleotide translocase (ANT) is a family of proteins involved in cell death pathways that perform distinctly opposite functions to regulate cell fate decisions. The results of this study indicate that under conditions found in the ischemic heart cell (low ATP, high phosphate), adenine nucleotides may be lost from the mitochondria via the adenine nucleotide translocase. T2 - State 3 respiration, adenine nucleotide translocase activity, and the net accumulation of adenine nucleotides. Cycles between the cytoplasmic-open state (c-state) and the matrix-open state (m-state): operates by the alternating access mechanism with a single substrate-binding site intermittently exposed to either the cytosolic (c-state) or matrix (m . Adenine nucleotide translocase-2 (ANT2) is an oncogenic m … EGFR tyrosine kinase inhibitor (EGFR-TKI) therapy has achieved favorable clinical outcomes in non-small cell lung cancer (NSCLC) patients with EGFR mutations. Molecular Weight: 770.82. Adenine nucleotide translocase was labeled as previously reported by Majima et al. In a minority of cases, it is associated with skeletal myopathy, which predominantly involves axial or proximal muscles and which causes abnormal fatigability and even permanent muscle weakness. T2 - State 3 respiration, adenine nucleotide translocase activity, and the net accumulation of adenine nucleotides. Targeting adenine nucleotide translocase-2 (ANT2) to overcome resistance to epidermal growth factor receptor tyrosine kinase inhibitor in non-small cell lung cancer. The adenine nucleotide translocase (ANT) is a dimeric protein complex of two identical 32 kDa subunits and facilitates the transport of ADP and ATP across the inner mitochondrial membrane . and membrane potential modulate the mitochondrial permeability transition by affecting nucleotide binding to the adenine nucleotide translocase. Polyporus officinalis and Polyporus igniarius. Abstract Adenine nucleotide translocase (AdNT) activity was studied in isolated mitochondria from normal rabbit aortas. J Biol Chem 279, 38415-38423. EGFR tyrosine kinase inhibitor (EGFR-TKI) therapy has achieved favorable clinical outcomes in non-small cell lung cancer (NSCLC) patients with EGFR mutations. . ANT is the most abundant protein in the inner mitochondrial membrane and belongs to mitochondrial carrier family.. Free ADP is transported from the cytoplasm to the . FEBS Lett 563, 155-160. Full text Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Current medicinal chemistry, 10(16), 1507-1525 (2003-07-23) Mitochondrial membrane permeabilization (MMP) is a rate-limiting step of apoptosis, including in anticancer chemotherapy. This thesis research aims to investigate the functional roles of TSPO and ANT2 in breast cancer progression and assess their Mitochondrial membrane carriers containing proline and cysteine, such as adenine nucleotide translocase (ANT), are potential targets of cyclophilin D (CyP-D) and potential Ca2+-induced permeability transition pore (PTP) components or regulators; Disease description A disorder characterized by progressive weakness of ocular muscles and levator muscle of the upper eyelid. In contrast, the second family, which includes bongkrekic acid (BA) and . the adenine nucleotide translocase, IMAC appears to be a separate entity, since some of the IC,, values differ by up to &fold, and carboxyatractyloside, the most selec- tive inhibitor of the adenine nucleotide translocase, has no effect on IMAC. In vitro the ADP/ATP . Andrew P Halestrap et al. The enzyme was inhibited by oleic acid, oleoylCoA, and oleoylcarnitine with 50% inhibition occurring at 5 muM, 6 muM and 14 muM, respectively (corresponding to 8, 10, and 23 nmol/mg protein). Specific mis-sense mutations in the human Ant1 . carboxyatractyloside is also inhibited (Table 2). Both compounds are currently being trialled in patients with solid tumours. 1. adenosine nucleotide translocase: antiproton, transports ADP3- into matrix in exchange for ATP4-, inhibited by atractyloside 2. phosphate translocase: symposia, one H2po4- and one H+ into matrix, favoured by transmembrane proton gradient In pathological conditions, F0F1-ATPase hydrolyzes ATP in an attempt to maintain mitochondrial membrane potential. Kinetic studies revealed atRA as an uncompetitive inhibitor of ANT. The mitochondrial membrane proteins, translocator protein (TSPO) and adenine nucleotide translocase 2 (ANT2), are both considered to be components of PTP complex and are overexpressed in cancers compared to normal tissues. Adenine nucleotide translocase-1 (ANT1) is an ADP/ATP transporter protein located in the inner mitochondrial membrane. Furthermore, by applying flux control theory to these inhibitor titrations, it was possible to demonstrate that the adenine nucleotide translocase exerted greater control over respiration in the newborn than in the adult, and at maximal rates of coupled respiration the translocase had a control strength of 0.98. Transient overexpression lysate of solute carrier family 25 (mitochondrial carrier; adenine nucleotide translocator), member 5 (SLC25A5), nuclear gene encoding mitochondrial protein The lysate was created in HEK293T cells, using Plasmid ID RC208949 and based on accession number NM_001152. specific translocase for ADP and ATP in the islet mitochondria (fig.1). Cancer Ther. Add a protease inhibitor cocktail to the mixture and keep the sample on ice until immunoprecipitation is performed. We conclude that agaric acid promotes the opening of the pore, increasing ROS production that exerts oxidative modification of critical thiols in the adenine . 268:153-160. Phenyl-succinate (20 mM) totally inhibited phosphate-induced efflux. In addition 50 µM quercetin inhibited the adenine nucleotide translocase (ANT) by 46%. Adenine nucleotide translocase (ANT) mediates the exchange of ADP and ATP on the inner mitochondrial membrane in healthy cells. Adenine nucleotide translocase in islet mitochondria. Adenine nucleotide translocase 3 (ANT3) overexpression induces apoptosis in cultured cells. 1997. ANT is the most abundant protein in the inner mitochondrial membrane and belongs to mitochondrial carrier family.. Free ADP is transported from the cytoplasm to the . Here, we describe the isolation of adenine nucleotide translocase-1 (ANT-1) in a screen for dominant, apoptosis-inducing genes. The mechanism of functional coupling between mitochondrial creatine kinase (MiCK) and adenine nucleotide translocase (ANT) in isolated heart mitochondria is analyzed. This is important, because it has long been assumed that the ADP/ATP translocase is the only adenine nucleotide transporter in mitochon-dna. 6C). Photoaffinity labeling of mitochondrial proteins with [3 H]atRA demonstrated the binding of a 31-kDa protein to atRA. In addition, ANT2 repression results in the growth arrest of human cells, implying that ANT2 is a candidate for cancer therapy based on molecular targeting. Adenine nucleotide translocase activity is known to be indiscriminately inhibited by a natural poison atractyloside (ATR) 5; 8; 23. Knockdown of ANT3 markedly inhibited the apoptotic induction . ANT-1 is a component of the mitochondrial permeability transition complex, a protein aggregate connecting the inner with the outer mitochondrial membrane that has recently been implicated in apoptosis. We demonstrated for the first time that retinoic acids inhibit adenine nucleotide translocase (ANT) activity in heart and liver mitochondria. In contrast to MDBNP, the average IC 50 values of ATR exhibited no statistical difference among ANT isoforms (Fig. 2005), while in primary human adipocytes, it inhibited adipogenesis even at 6.25 . We showed that CPT-induced apoptosis in QGY7703 cells and down-regulated the expression of ANT3. Halestrap's group (21-23) has suggested that the adenine nucleotide translocase may be responsible for K+ uniport and that cyclophilin is an important regulator. Here, we describe the isolation of adenine nucleotide translocase-1 (ANT-1) in a screen for dominant, apoptosis-inducing genes. The results of this study indicate that under conditions found in the ischemic heart cell (low ATP, high phosphate), adenine nucleotides may be lost from the mitochondria via the adenine nucleotide translocase. This protein was . Adenine nucleotide translocator (ANT), also known as the ADP/ATP translocase (ANT), ADP/ATP carrier protein (AAC) or mitochondrial ADP/ATP carrier, exchanges free ATP with free ADP across the inner mitochondrial membrane. Bcl-2-family proteins. A new endogenous ATP analog (ApppI) inhibits the mitochondrial adenine nucleotide translocase (ANT) and is responsible for the apoptosis induced by nitrogen-containing bisphosphonates. ANT1, ANT2 and ANT3 paralogs are expressed in normal human bronchial epithelial cells with ANT2 being predominant . ; 1 mg of mitochondrial protein was suspended in 1 ml of a basic medium and preincubated with 3 μM agaric acid and 20 μM tamoxifen for 5 min; subsequently, 20 nmol/mg eosin maleimide (EMA) was added and incubated for 5 . Hannu Mönkkönen, . Adenine Nucleotide Translocase Immunocapture Kit Instructions for Use For the isolation of Adenine Nucleotide . Agaricic acid is an adenine nucleotide translocase antagonist.It is obtained from various plants of the fungous tribe, i.e. ANT is the most abundant protein in the inner mitochondrial membrane and belongs to mitochondrial carrier family. The human genome encodes four paralogs (ANT1-4), with variable tissue expression (Liu and Chen, 2013). The kinetic properties and inhibitor sensi- . ApppI inhibits mitochondrial adenine nucleotide translocase (ANT) HMG-CoA reductase) and zoledronic acid. Phenyl-succinate (20 mM) totally inhibited phosphate-induced efflux. ANT therefore plays an important role in cellular energy metabolism by influencing mitochondrial oxidative phosphorylation. Mersalyl (10 nmol/mg protein) did not inhibit efflux. Regulation of the mitochondrial adenine nucleotide pooi size in liver: mechanism . The effect of agaric acid was inhibited by the antioxidant tamoxifen in association with decreased binding of the thiol reagent eosin-3 maleimide to the adenine nucleotide translocase. . At 0.2-2 mM To further analyze the molecular mechanisms of action of concentrations, lovastatin inhibited the IPP accumulation and ApppI, we next studied its effect in the isolated rat liver the ApppI production (Po0.001) in a dose . Recruitment of NF-kappaB into mitochondria is involved in adenine nucleotide translocase 1 (ANT1)-induced apoptosis. Translocase was labeled as previously reported by Majima et al apoptosis has not been examined a! 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