N-Glycosylation of the Na+-Taurocholate Cotransporting Polypeptide (NTCP) Determines Its Trafficking and Stability and Is Required for Hepatitis B Virus Infection

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Jan 27

PMID 28125599

Citations 23

Authors

Monique D Appelman

Anindita Chakraborty

Ulrike Protzer

Jane A McKeating

Stan F J van de Graaf

Affiliations

Soon will be listed here.

Abstract

The sodium/bile acid cotransporter NTCP was recently identified as a receptor for hepatitis B virus (HBV). NTCP is glycosylated and the role of glycans in protein trafficking or viral receptor activity is not known. NTCP contains two N-linked glycosylation sites and asparagine amino acid residues N5 and N11 were mutated to a glutamine to generate NTCP with a single glycan (NTCP-N5Q or NTCP- N11Q) or no glycans (NTCP- N5,11Q). HepG2 cells expressing NTCP with a single glycan supported HBV infection at a comparable level to NTCP-WT. The physiological function of NTCP, the uptake of bile acids, was also not affected in cells expressing these single glycosylation variants, consistent with their trafficking to the plasma membrane. However, glycosylation-deficient NTCP (NTCP-N5,11Q) failed to support HBV infection, showed minimal cellular expression and was degraded in the lysosome. This affected the physiological bile acid transporter function of NTCP-N5,11Q in a similar fashion. In conclusion, N-glycosylation is required for efficient NTCP localization at the plasma membrane and subsequent HBV infection and these characteristics are preserved in NTCP carrying a single carbohydrate moiety.

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References

Vaz F, Paulusma C, Huidekoper H, de Ru M, Lim C, Koster J . Sodium taurocholate cotransporting polypeptide (SLC10A1) deficiency: conjugated hypercholanemia without a clear clinical phenotype. Hepatology. 2014; 61(1):260-7. DOI: 10.1002/hep.27240. View

Console L, Scalise M, Tarmakova Z, Coe I, Indiveri C . N-linked glycosylation of human SLC1A5 (ASCT2) transporter is critical for trafficking to membrane. Biochim Biophys Acta. 2015; 1853(7):1636-45. DOI: 10.1016/j.bbamcr.2015.03.017. View

Schulze A, Gripon P, Urban S . Hepatitis B virus infection initiates with a large surface protein-dependent binding to heparan sulfate proteoglycans. Hepatology. 2007; 46(6):1759-68. DOI: 10.1002/hep.21896. View

Meredith L, Hu K, Cheng X, Howard C, Baumert T, Balfe P . Lentiviral hepatitis B pseudotype entry requires sodium taurocholate co-transporting polypeptide and additional hepatocyte-specific factors. J Gen Virol. 2015; 97(1):121-127. PMC: 4772705. DOI: 10.1099/jgv.0.000317. View

Sun A, Swaby I, Xu S, Suchy F . Cell-specific basolateral membrane sorting of the human liver Na(+)-dependent bile acid cotransporter. Am J Physiol Gastrointest Liver Physiol. 2001; 280(6):G1305-13. DOI: 10.1152/ajpgi.2001.280.6.G1305. View

Hagenbuch B, Meier P . Molecular cloning, chromosomal localization, and functional characterization of a human liver Na+/bile acid cotransporter. J Clin Invest. 1994; 93(3):1326-31. PMC: 294097. DOI: 10.1172/JCI117091. View

Blank A, Markert C, Hohmann N, Carls A, Mikus G, Lehr T . First-in-human application of the novel hepatitis B and hepatitis D virus entry inhibitor myrcludex B. J Hepatol. 2016; 65(3):483-9. DOI: 10.1016/j.jhep.2016.04.013. View

Yan H, Peng B, Liu Y, Xu G, He W, Ren B . Viral entry of hepatitis B and D viruses and bile salts transportation share common molecular determinants on sodium taurocholate cotransporting polypeptide. J Virol. 2014; 88(6):3273-84. PMC: 3957944. DOI: 10.1128/JVI.03478-13. View

Slijepcevic D, Kaufman C, Wichers C, Gilglioni E, Lempp F, Duijst S . Impaired uptake of conjugated bile acids and hepatitis b virus pres1-binding in na(+) -taurocholate cotransporting polypeptide knockout mice. Hepatology. 2015; 62(1):207-19. PMC: 4657468. DOI: 10.1002/hep.27694. View

10.

Yan H, Zhong G, Xu G, He W, Jing Z, Gao Z . Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. Elife. 2012; 1:e00049. PMC: 3485615. DOI: 10.7554/eLife.00049. View

11.

CRADDOCK A, Love M, Daniel R, Kirby L, Walters H, Wong M . Expression and transport properties of the human ileal and renal sodium-dependent bile acid transporter. Am J Physiol. 1998; 274(1):G157-69. DOI: 10.1152/ajpgi.1998.274.1.G157. View

12.

Pereira I, Buchmann B, Sandmann L, Sprinzl K, Schlaphoff V, Dohner K . Primary biliary acids inhibit hepatitis D virus (HDV) entry into human hepatoma cells expressing the sodium-taurocholate cotransporting polypeptide (NTCP). PLoS One. 2015; 10(2):e0117152. PMC: 4315608. DOI: 10.1371/journal.pone.0117152. View

13.

Zhao N, Zhang A, Worthen C, Knutson M, Enns C . An iron-regulated and glycosylation-dependent proteasomal degradation pathway for the plasma membrane metal transporter ZIP14. Proc Natl Acad Sci U S A. 2014; 111(25):9175-80. PMC: 4078863. DOI: 10.1073/pnas.1405355111. View

14.

Beers M, Zhao M, Tomer Y, Russo S, Zhang P, Gonzales L . Disruption of N-linked glycosylation promotes proteasomal degradation of the human ATP-binding cassette transporter ABCA3. Am J Physiol Lung Cell Mol Physiol. 2013; 305(12):L970-80. PMC: 3882536. DOI: 10.1152/ajplung.00184.2013. View

15.

Protzer U, Seyfried S, Quasdorff M, Sass G, Svorcova M, Webb D . Antiviral activity and hepatoprotection by heme oxygenase-1 in hepatitis B virus infection. Gastroenterology. 2007; 133(4):1156-65. DOI: 10.1053/j.gastro.2007.07.021. View

16.

Jost S, Turelli P, Mangeat B, Protzer U, Trono D . Induction of antiviral cytidine deaminases does not explain the inhibition of hepatitis B virus replication by interferons. J Virol. 2007; 81(19):10588-96. PMC: 2045472. DOI: 10.1128/JVI.02489-06. View

17.

Yoshimori T, Yamamoto A, Moriyama Y, Futai M, Tashiro Y . Bafilomycin A1, a specific inhibitor of vacuolar-type H(+)-ATPase, inhibits acidification and protein degradation in lysosomes of cultured cells. J Biol Chem. 1991; 266(26):17707-12. View

18.

Verrier E, Colpitts C, Bach C, Heydmann L, Weiss A, Renaud M . A targeted functional RNA interference screen uncovers glypican 5 as an entry factor for hepatitis B and D viruses. Hepatology. 2015; 63(1):35-48. DOI: 10.1002/hep.28013. View

19.

Petersen J, Dandri M, Mier W, Lutgehetmann M, Volz T, von Weizsacker F . Prevention of hepatitis B virus infection in vivo by entry inhibitors derived from the large envelope protein. Nat Biotechnol. 2008; 26(3):335-41. DOI: 10.1038/nbt1389. View

20.

Ni Y, Lempp F, Mehrle S, Nkongolo S, Kaufman C, Falth M . Hepatitis B and D viruses exploit sodium taurocholate co-transporting polypeptide for species-specific entry into hepatocytes. Gastroenterology. 2013; 146(4):1070-83. DOI: 10.1053/j.gastro.2013.12.024. View