Transportomics: screening for substrates of ABC transporters in body fluids using vesicular transport assays

Krumpochova, Petra; Sapthu, Sunny; Brouwers, Jos F.; Haas, Marcel de; Vos, Ric C. H. de; Borst, Piet; Wetering, Koen van de

doi:10.1096/fj.11-195743

Cited by 54 publications

(56 citation statements)

References 27 publications

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“…Conjugation of enterolignans with sulfate and glucuronic acid occurs in the intestinal wall and liver, with the predominant conjugates being glucuronides. Both sulfate and glucuronide conjugates have been identified as ABCC2 and ABCG2 substrates (Lampe et al, 2006;Krumpochova et al, 2012;van de Wetering and Sapthu, 2012). These authors found that ABCG2 was able to transport the enterolactone sulfate with low affinity, and its disposition in Abcg2 2/2 mice was altered, highlighting the physiologic relevance of this result.…”

Section: Resultsmentioning

confidence: 99%

Role of ABCG2 in Transport of the Mammalian Lignan Enterolactone and its Secretion into Milk in Abcg2 Knockout Mice

Miguel

Otero

Garcı́a-Villalba

et al. 2014

Drug Metabolism and Disposition

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Lignans are phytoestrogens that are metabolized by the gut microbiota to enterodiol and enterolactone, the main biologically active enterolignans. Substantial interindividual variation in plasma concentration and urinary excretion of enterolignans has been reported, this being determined, at least in part, by the intake of lignan precursors, the gut microbiota, and the host's phase 2 conjugating enzyme activity. However, the role of ATP-binding cassette (ABC) transporters in the transport and disposition of enterolactone has not been reported so far. Active transport assays using parental and Madin-Darby canine kidney epithelial cells transduced with murine and human ABCG2 showed a significant increase in apically directed translocation of enterolactone in transduced cells, which was confirmed by using the selective ABCG2 inhibitor Ko143. In addition, enterolactone also inhibited transport of the antineoplastic agent mitoxantrone as a model substrate, with inhibition percentages of almost 40% at 200 mM for human ABCG2. Furthermore, the endogenous levels in plasma and milk of enterolactone in wild-type and Abcg2(2/2) knockout female mice were analyzed. The milk/ plasma ratio decreased significantly in the Abcg2 (2/2) phenotype, as compared with the wild-type mouse group (0.4 6 0.1 as against 6.4 6 2.6). This paper is the first to report that enterolactone is a transported substrate and therefore most probably a competitive inhibitor of ABCG2, which suggests it has a role in the interindividual variations in the disposition of enterolactone and its secretion into milk. The inhibitory activity identified provides a solid basis for further investigation in possible food-drug interactions. IntroductionThe ATP-binding cassette (ABC) subfamily G2 (ABCG2) protein is apically expressed ABC membrane transporter that mediates the active and outward transport of a wide range of anticancer drugs, dietary compounds, food carcinogens, and antibiotics from cells (van Herwaarden et al., 2003;Merino et al., 2005;Vlaming et al., 2009). In recent years, its role as a transporter of phytoestrogens and their conjugated metabolites has gained special relevance (Zhu et al., 2010;Alvarez et al., 2011;Tan et al., 2013).ABCG2 is found in tumor cells, but also in the cells of a variety of normal tissues involved in the uptake and elimination of compounds, such as enterocytes, hepatocytes, and cells of the proximal tubules of the kidney. The apical localization in these cells allows ABCG2 to mediate hepatobiliary and urinary elimination and to function as a barrier to uptake from the gut lumen (van Herwaarden and Schinkel, 2006). In addition, induced expression of ABCG2 in the lactating mammary gland supports its important role in the active secretion of several xenobiotics and beneficial compounds such as vitamins into milk Merino et al., 2006;van Herwaarden et al., 2007).Dietary intake of lignans and their derivatives, as part of a healthy diet, has been associated with protective effects against a number of chronic diseases (Hu e...

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Section: Resultsmentioning

confidence: 99%

Role of ABCG2 in Transport of the Mammalian Lignan Enterolactone and its Secretion into Milk in Abcg2 Knockout Mice

Miguel

Otero

Garcı́a-Villalba

et al. 2014

Drug Metabolism and Disposition

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“…Its endogenous substrates include tetrahydroxylated bile acids (Megaraj et al, 2010), divalent bile acids dmd.aspetjournals.org (Kuipers et al, 1988), glutathione (Oude Elferink et al, 1990), bilirubin glucuronosides (Paulusma et al, 1997), eicosanoids (prostaglandin E2, leukotriene C 4 ) , and conjugated steroids [estrone 3-sulfate (Kopplow et al, 2005), estradiol-17b-glucuronate ]. Exogenous ABCC2 substrates are mostly conjugated, either with glucuronic acid [e.g., phytoestrogens (Krumpochova et al, 2012), acetaminophen (Xiong et al, 2000), indomethacin (Kouzuki et al, 2000), morphine (van de Wetering et al, 2007)], sulfuric acid [e.g., acetaminophen (Zamek-Gliszczynski et al, 2005), resveratrol (Kaldas et al, 2003)], or with glutathione [e.g., acetaminophen (Chen et al, 2003a), bromosulfophthalein (Jansen et al, 1987), dinitrophenyl (Elferink et al, 1989)]. However, ABCC2 also transports unconjugated anionic drugs, such as pravastatin (Yamazaki et al, 1997), ampicillin (Verkade et al, 1990), and methotrexate (Hooijberg et al, 1999).…”

Section: Abcc2mentioning

confidence: 99%

The Role of Canalicular ABC Transporters in Cholestasis

Cuperus

Claudel

Gautherot

et al. 2014

Drug Metabolism and Disposition

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Cholestasis, a hallmark feature of hepatobiliary disease, is characterized by the retention of biliary constituents. Some of these constituents, such as bile acids, inflict damage to hepatocytes and bile duct cells. This damage may lead to inflammation, fibrosis, cirrhosis, and eventually carcinogenesis, sequelae that aggravate the underlying disease and deteriorate clinical outcome. Canalicular ATP-binding cassette (ABC) transporters, which mediate the excretion of individual bile constituents, play a key role in bile formation and cholestasis. The study of these transporters and their regulatory nuclear receptors has revolutionized our understanding of cholestatic disease. This knowledge has served as a template to develop novel treatment strategies, some of which are currently already undergoing phase III clinical trials. In this review we aim to provide an overview of the structure, function, and regulation of canalicular ABC transporters. In addition, we will focus on the role of these transporters in the pathogenesis and treatment of cholestatic bile duct and liver diseases.

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“…In previous experiments, we have used semitargeted metabolomics to discover endogenous substrates of ABCC transporters (26,27). Because this approach requires some prior knowledge of potential substrates, it is not applicable to ABCC5.…”

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confidence: 99%

N -lactoyl-amino acids are ubiquitous metabolites that originate from CNDP2-mediated reverse proteolysis of lactate and amino acids

Jansen

Addie

Merkx³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Despite technological advances in metabolomics, large parts of the human metabolome are still unexplored. In an untargeted metabolomics screen aiming to identify substrates of the orphan transporter ATP-binding cassette subfamily C member 5 (ABCC5), we identified a class of mammalian metabolites, N-lactoyl-amino acids. Using parallel protein fractionation in conjunction with shotgun proteomics on fractions containing N-lactoyl-Phe-forming activity, we unexpectedly found that a protease, cytosolic nonspecific dipeptidase 2 (CNDP2), catalyzes their formation. N-lactoyl-amino acids are ubiquitous pseudodipeptides of lactic acid and amino acids that are rapidly formed by reverse proteolysis, a process previously considered to be negligible in vivo. The plasma levels of these metabolites strongly correlate with plasma levels of lactate and amino acid, as shown by increased levels after physical exercise and in patients with phenylketonuria who suffer from elevated Phe levels. Our approach to identify unknown metabolites and their biosynthesis has general applicability in the further exploration of the human metabolome.unknown metabolites | untargeted metabolomics | ABCC5 | MRP5 | physical exercise U ntargeted metabolomics aims to provide a comprehensive snapshot of the metabolome and is becoming a mainstream technique to discover biomarkers, to study the effects of interventions, and to discover the function of enzymes (1).Recent technical improvements now make it possible to detect several thousand metabolites in a single untargeted metabolomics analysis, but the identity of these metabolites is initially not known beyond their molecular mass (1). Knowing the chemical identity of metabolites is crucial for the proper interpretation of metabolomic studies, however. Online metabolite databases like METLIN (∼240,000 entries) and the Human Metabolome Database (HMDB; ∼42,000 entries) contain vast numbers of metabolites and are extremely useful to annotate the detected molecular masses (2, 3). These databases cover large parts of the metabolome, but significant gaps remain. Due to poorly characterized and promiscuous enzymes, the human metabolome is much larger than initially anticipated (4). State-of-the-art untargeted metabolomics studies still report up to 40% unidentified, but potentially important, metabolites that can be detected reproducibly (5-8). Nevertheless, unknown metabolites are only rarely characterized because of the extensive work required for de novo structure elucidation (9-11). Here, we describe the characterization of a class of mammalian metabolites that we discovered in an untargeted metabolomic screen aimed to identify substrates of the orphan transporter ATP-binding cassette subfamily C member 5 (ABCC5).ABCC5 is a member of the C-branch of the superfamily of ABC transporters, which use the energy provided by the hydrolysis of ATP to transport substrates across the plasma membrane (12). ABCC5, also known as multidrug resistance-associated protein 5 (MRP5), is ubiquitously expressed, but levels in b...

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Transportomics: screening for substrates of ABC transporters in body fluids using vesicular transport assays

Cited by 54 publications

References 27 publications

Role of ABCG2 in Transport of the Mammalian Lignan Enterolactone and its Secretion into Milk in Abcg2 Knockout Mice

Role of ABCG2 in Transport of the Mammalian Lignan Enterolactone and its Secretion into Milk in Abcg2 Knockout Mice

The Role of Canalicular ABC Transporters in Cholestasis

N -lactoyl-amino acids are ubiquitous metabolites that originate from CNDP2-mediated reverse proteolysis of lactate and amino acids

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