Skip to main content Accessibility help

Effect of bovine ABCG2 polymorphism Y581S SNP on secretion into milk of enterolactone, riboflavin and uric acid

  • J. A. Otero (a1) (a2), V. Miguel (a1) (a2), L. González-Lobato (a2), R. García-Villalba (a3), J. C. Espín (a3), J. G. Prieto (a2) (a4), G. Merino (a1) (a2) and A. I. Álvarez (a1) (a2)...


The ATP-binding cassette transporter G2/breast cancer resistance protein (ABCG2/BCRP) is an efflux protein involved in the bioavailability and milk secretion of endogenous and exogenous compounds, actively affecting milk composition. A limited number of physiological substrates have been identified. However, no studies have reported the specific effect of this polymorphism on the secretion into milk of compounds implicated in milk quality such as vitamins or endogenous compounds. The bovine ABCG2 Y581S polymorphism is described as a gain-of-function polymorphism that increases milk secretion and decreases plasma levels of its substrates. This work aims to study the impact of Y581S polymorphism on plasma disposition and milk secretion of compounds such as riboflavin (vitamin B2), enterolactone, a microbiota-derived metabolite from the dietary lignan secoisolariciresinol and uric acid. In vitro transport of these compounds was assessed in MDCK-II cells overexpressing the bovine ABCG2 (WT-bABCG2) and its Y581S variant (Y581S-bABCG2). Plasma and milk levels were obtained from Y/Y homozygous and Y/S heterozygous cows. The results show that riboflavin was more efficiently transported in vitro by the Y581S variant, although no differences were noted in vivo. Both uric acid and enterolactone were substrates in vitro of the bovine ABCG2 variants and were actively secreted into milk with a two-fold increase in the milk/plasma ratio for Y/S with respect to Y/Y cows. The in vitro ABCG2-mediated transport of the drug mitoxantrone, as a model substrate, was inhibited by enterolactone in both variants, suggesting the possible in vivo use of this enterolignan to reduce ABCG2-mediated milk drug transfer in cows. The Y581S variant was inhibited to a lesser extent probably due to its higher transport capacity. All these findings point to a significant role of the ABCG2 Y581S polymorphism in the milk disposition of enterolactone and the endogenous molecules riboflavin and uric acid, which could affect both milk quality and functionality.


Corresponding author



Hide All
Adolphe, JL, Whiting, SJ, Juurlink, BH, Thorpe, LU and Alcorn, J 2010. Health effects with consumption of the flax lignan secoisolariciresinol diglucoside. The British Journal of Nutrition 103, 929938.
Alvarez, AI, Real, R, Perez, M, Mendoza, G, Prieto, JG and Merino, G 2010. Modulation of the activity of ABC transporters (P-glycoprotein, MRP2, BCRP) by flavonoids and drug response. Journal of Pharmaceutical Sciences 99, 598617.
Antignac, J, Cariou, R, Le-Bizec, B and André, F 2004. New data regarding phytoestrogens content in bovine milk. Food Chemistry 87, 275281.
Aycicek, A, Erel, O, Kocyigit, A, Selek, S and Demirkol, MR 2006. Breast milk provides better antioxidant power than does formula. Nutrition (Burbank, Los Angeles County, Calif.) 22, 616619.
Bolca, S, Urpi-Sarda, M, Blondeel, P, Roche, N, Vanhaecke, L, Possemiers, S, Al-Maharik, N, Botting, N, De Keukeleire, D, Bracke, M, Heyerick, A, Manach, C and Depypere, H 2010. Disposition of soy isoflavones in normal human breast tissue. The American Journal of Clinical Nutrition 91, 976984.
Cohen-Zinder, M, Seroussi, E, Larkin, DM, Loor, JJ, Everts-van der Wind, A, Lee, JH, Drackley, JK, Band, MR, Hernandez, AG, Shani, M, Lewin, HA, Weller, JI and Ron, M 2005. Identification of a missense mutation in the bovine ABCG2 gene with a major effect on the QTL on chromosome 6 affecting milk yield and composition in Holstein cattle. Genome Research 15, 936944.
Cortes, C, Palin, MF, Gagnon, N, Benchaar, C, Lacasse, P and Petit, HV 2012. Mammary gene expression and activity of antioxidant enzymes and concentration of the mammalian lignan enterolactone in milk and plasma of dairy cows fed flax lignans and infused with flax oil in the abomasum. The British Journal of Nutrition 108, 13901398.
Dankers, AC, Mutsaers, HA, Dijkman, HB, van den Heuvel, LP, Hoenderop, JG, Sweep, FC, Russel, FG and Masereeuw, R 2013. Hyperuricemia influences tryptophan metabolism via inhibition of multidrug resistance protein 4 (MRP4) and breast cancer resistance protein (BCRP). Biochimica et Biophysica Acta 1832, 17151722.
Gagnon, N, Cortes, C, da Silva, D, Kazama, R, Benchaar, C, dos Santos, G, Zeoula, L and Petit, HV 2009. Ruminal metabolism of flaxseed (Linum usitatissimum) lignans to the mammalian lignan enterolactone and its concentration in ruminal fluid, plasma, urine and milk of dairy cows. The British Journal of Nutrition 102, 10151023.
Gonzalez-Lobato, L, Real, R, Herrero, D, de la Fuente, A, Prieto, JG, Marques, MM, Alvarez, AI and Merino, G 2014. Novel in vitro systems for prediction of veterinary drug residues in ovine milk and dairy products. Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment 31, 10261037.
Haslam, IS and Simmons, NL 2014. Expression of the ABC transport proteins MDR1 (ABCB1) and BCRP (ABCG2) in bovine rumen. Journal of Comparative Physiology. B, Biochemical, Systemic, and Environmental Physiology 184, 673681.
Hosomi, A, Nakanishi, T, Fujita, T and Tamai, I 2012. Extra-renal elimination of uric acid via intestinal efflux transporter BCRP/ABCG2. PLoS One 7, e30456.
Jani, M and Krajcsi, P 2014. In vitro methods in drug transporter interaction assessment. Drug Discovery Today Technologies 12, e105e112.
Jonker, JW, Merino, G, Musters, S, van Herwaarden, AE, Bolscher, E, Wagenaar, E, Mesman, E, Dale, TC and Schinkel, AH 2005. The breast cancer resistance protein BCRP (ABCG2) concentrates drugs and carcinogenic xenotoxins into milk. Nature Medicine 11, 127129.
Koop, J, Monschein, S, Pauline Macheroux, E, Knaus, T and Macheroux, P 2014. Determination of free and bound riboflavin in cow's milk using a novel flavin-binding protein. Food Chemistry 146, 9497.
Larsen, T and Moyes, KM 2010. Fluorometric determination of uric acid in bovine milk. The Journal of Dairy Research 77, 438444.
Lindner, S, Halwachs, S, Wassermann, L and Honscha, W 2013. Expression and subcellular localization of efflux transporter ABCG2/BCRP in important tissue barriers of lactating dairy cows, sheep and goats. Journal of Veterinary Pharmacology and Therapeutics 36, 562570.
Liu, X, Lin, WM, Yan, XH, Chen, XH, Hoidal, JR and Xu, P 2003. Improved method for measurement of human plasma xanthine oxidoreductase activity. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences 785, 101114.
Mealey, KL 2012. ABCG2 transporter: therapeutic and physiologic implications in veterinary species. Journal of Veterinary Pharmacology and Therapeutics 35, 105112.
Merino, G, Real, R, Baro, MF, Gonzalez-Lobato, L, Prieto, JG, Alvarez, AI and Marques, MM 2009. Natural allelic variants of bovine ATP-binding cassette transporter ABCG2: increased activity of the Ser581 variant and development of tools for the discovery of new ABCG2 inhibitors. Drug Metabolism and Disposition: The Biological Fate of Chemicals 37, 59.
Miguel, V, Otero, JA, Garcia-Villalba, R, Tomas-Barberan, F, Espin, JC, Merino, G and Alvarez, AI 2014. Role of ABCG2 in transport of the mammalian lignan enterolactone and its secretion into milk in Abcg2 knockout mice. Drug Metabolism and Disposition: The Biological Fate of Chemicals 42, 943946.
Njastad, KM, Adler, SA, Hansen-Moller, J, Thuen, E, Gustavsson, AM and Steinshamn, H. 2014. Gastrointestinal metabolism of phytoestrogens in lactating dairy cows fed silages with different botanical composition. Journal of Dairy Science 97, 77357750.
Olsen, HG, Nilsen, H, Hayes, B, Berg, PR, Svendsen, M, Lien, S and Meuwissen, T 2007. Genetic support for a quantitative trait nucleotide in the ABCG2 gene affecting milk composition of dairy cattle. BMC Genetics 8, 32.
Otero, JA, Barrera, B, de la Fuente, A, Prieto, JG, Marques, M, Alvarez, AI and Merino, G 2015. Short communication: The gain-of-function Y581S polymorphism of the ABCG2 transporter increases secretion into milk of danofloxacin at the therapeutic dose for mastitis treatment. Journal of Dairy Science 98, 312317.
Otero, JA, Real, R, de la Fuente, A, Prieto, JG, Marques, M, Alvarez, AI and Merino, G 2013. The bovine ATP-binding cassette transporter ABCG2 Tyr581Ser single-nucleotide polymorphism increases milk secretion of the fluoroquinolone danofloxacin. Drug Metabolism and Disposition: The Biological Fate of Chemicals 41, 546549.
Pavek, P, Merino, G, Wagenaar, E, Bolscher, E, Novotna, M, Jonker, JW and Schinkel, AH 2005. Human breast cancer resistance protein: interactions with steroid drugs, hormones, the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine, and transport of cimetidine. The Journal of Pharmacology and Experimental Therapeutics 312, 144152.
Petit, HV, Gagnon, N, Mir, PS, Cao, R and Cui, S 2009. Milk concentration of the mammalian lignan enterolactone, milk production, milk fatty acid profile, and digestibility in dairy cows fed diets containing whole flaxseed or flaxseed meal. The Journal of Dairy Research 76, 257264.
Pulido, MM, Molina, AJ, Merino, G, Mendoza, G, Prieto, JG and Alvarez, AI 2006. Interaction of enrofloxacin with breast cancer resistance protein (BCRP/ABCG2): influence of flavonoids and role in milk secretion in sheep. Journal of Veterinary Pharmacology and Therapeutics 29, 279287.
Real, R, Gonzalez-Lobato, L, Baro, MF, Valbuena, S, de la Fuente, A, Prieto, JG, Alvarez, AI, Marques, MM and Merino, G 2011. Analysis of the effect of the bovine adenosine triphosphate-binding cassette transporter G2 single nucleotide polymorphism Y581S on transcellular transport of veterinary drugs using new cell culture models. Journal of Animal Science 89, 432543238.
Ron, M, Cohen-Zinder, M, Peter, C, Weller, JI and Erhardt, G 2006. Short communication: a polymorphism in ABCG2 in Bos indicus and Bos taurus cattle breeds. Journal of Dairy Science 89, 49214923.
Safranow, K, Machoy, Z and Ciechanowski, K 2000. Analysis of purines in urinary calculi by high-performance liquid chromatography. Analytical Biochemistry 286, 224230.
Schogor, AL, Palin, MF, Santos, GT, Benchaar, C, Lacasse, P and Petit, HV 2013. Mammary gene expression and activity of antioxidant enzymes and oxidative indicators in the blood, milk, mammary tissue and ruminal fluid of dairy cows fed flax meal. The British Journal of Nutrition 110, 17431750.
Steinshamn, H, Purup, S, Thuen, E and Hansen-Moller, J 2008. Effects of clover-grass silages and concentrate supplementation on the content of phytoestrogens in dairy cow milk. Journal of Dairy Science 91, 27152725.
van Herwaarden, AE, Wagenaar, E, Merino, G, Jonker, JW, Rosing, H, Beijnen, JH and Schinkel, AH 2007. Multidrug transporter ABCG2/breast cancer resistance protein secretes riboflavin (vitamin B2) into milk. Molecular and Cellular Biology 27, 12471253.
Wassermann, L, Halwachs, S, Lindner, S, Honscha, KU and Honscha, W 2013. Determination of functional ABCG2 activity and assessment of drug-ABCG2 interactions in dairy animals using a novel MDCKII in vitro model. Journal of Pharmaceutical Sciences 102, 772784.
Yonezawa, A and Inui, K 2013. Novel riboflavin transporter family RFVT/SLC52: identification, nomenclature, functional characterization and genetic diseases of RFVT/SLC52. Molecular Aspects of Medicine 34, 693701.



Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed