Advertisement

Identification of the multivalent PDZ protein PDZK1 as a binding partner of sodium–coupled monocarboxylate transporter SMCT1 (SLC5A8) and SMCT2 (SLC5A12)

  • Sunena Srivastava
  • Kiyoshi Nakagawa
  • Xin He
  • Toru Kimura
  • Toshiyuki Fukutomi
  • Seiji Miyauchi
  • Hiroyuki Sakurai
  • Naohiko Anzai
Original Paper
  • 50 Downloads

Abstract

Sodium-coupled monocarboxylate transporters SMCT1 (SLC5A8) and SMCT2 (SLC5A12) mediate the high- and low-affinity transport of lactate in the kidney, but their regulatory mechanism is still unknown. Since these two transporters have the PDZ-motif at their C-terminus, the function of SMCTs may be modulated by a protein–protein interaction. To investigate the binding partner(s) of SMCTs in the kidney, we performed yeast two-hybrid (Y2H) screenings of a human kidney cDNA library with the C-terminus of SMCT1 (SMCT1-CT) and SMCT2 (SMCT2-CT) as bait. PDZK1 was identified as a partner for SMCTs. PDZK1 coexpression in SMCT1-expressing HEK293 cells enhanced their nicotinate transport activity. PDZK1, SMCT1, and URAT1 in vitro assembled into a single tri-molecular complex and their colocalization was confirmed in the renal proximal tubule in vivo by immunohistochemistry. These results indicate that the SMCT1-PDZK1 interaction thus plays an important role in both lactate handling as well as urate reabsorption in the human kidney.

Keywords

Monocarboxylates Monocarboxylate transporter SMCT Lactate PDZ PDZK1 

Abbreviations

URAT1

Urate transporter 1

SLC

Solute carrier

SMCTs

Sodium–coupled monocarboxylate transporter

PDZ

PSD-95, DglA, and ZO-1

Notes

Acknowledgements

The authors thank A. Toki, R. Kofuji, A. Yamanishi, and H. Miyazaki for technical assistance. This work was supported in part by Grants from JSPS (KAKENHI 15590233, 18590900, 21390073, 26461258, 18K08200), the Salt Science Research Foundation (No. 0524, 0721), The Nakatomi Foundation, Gout Research Foundation of Japan, and Kyorin University School of Medicine (Collaborative Project 2009) to N.A. This work was presented in part at the Renal Week 2006 of the American Society of Nephrology, San Diego, CA, November 2006, and at the Annual Meeting of Experimental Biology 2008, San Diego, CA, April 2008.

Author contributions

NA designed the study. SS and NA performed the Y2H. NA, KN and SM checked and confirmed the experimental materials and data used in this study. SS and XH performed the transport assay. SS, TK and TF performed the biochemical and immunohistochemical analysis. SS, NA and HS wrote the manuscript. All authors discussed the results and commented on the manuscript.

Compliance with ethical standards

Conflict of interest

All authors of this manuscript declare that we have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. 1.
    Poole RC, Halestrap AP (1993) Transport of lactate and other monocarboxylates across mammalian plasma membranes. Am J Physiol 264:C761–C782CrossRefGoogle Scholar
  2. 2.
    Murer H, Manganel M, Roch-Ramel F (1992) Tubular transport of monocarboxylates, Krebs cycle intermediates, and inorganic sulfate. In: Windhager EE (ed) Handbook of physiology section 8: renal physiology. Oxford University Press, New YorkGoogle Scholar
  3. 3.
    Jorgensen KE, Sheikh MI (1984) Renal transport of monocarboxylic acids. Heterogeneity of lactate-transport systems along the proximal tubule. Biochem J 223:803–807CrossRefGoogle Scholar
  4. 4.
    Miyauchi S, Gopal E, Fei YJ, Ganapathy V (2004) Functional identification of SLC5A8, a tumor suppressor down-regulated in colon cancer, as a Na+-coupled transporter for short-chain fatty acids. J Biol Chem 279:13293–13296CrossRefGoogle Scholar
  5. 5.
    Gopal E, Fei YJ, Sugawara M, Miyauchi S, Zhuang L, Martin P, Smith SB, Prasad PD, Ganapathy V (2004) Expression of slc5a8 in kidney and its role in Na+-coupled transport of lactate. J Biol Chem 279:44522–44532CrossRefGoogle Scholar
  6. 6.
    Coady MJ, Chang MH, Charron FM, Plata C, Wallendorff B, Sah JF, Markowitz SD, Romero MF, Lapointe JY (2004) The human tumour suppressor gene SLC5A8 expresses a Na+-monocarboxylate cotransporter. J Physiol 557:719–731CrossRefGoogle Scholar
  7. 7.
    Gopal E, Fei YJ, Miyauchi S, Zhuang L, Prasad PD, Ganapathy V (2005) Sodium-coupled and electrogenic transport of B-complex vitamin nicotinic acid by slc5a8, a member of the Na+/glucose co-transporter gene family. Biochem J 388:309–316CrossRefGoogle Scholar
  8. 8.
    Srinivas SR, Gopal E, Zhuang L, Itagaki S, Martin PM, Fei YJ, Ganapathy V, Prasad PD (2005) Cloning and functional identification of slc5a12 as a sodium-coupled low-affinity transporter for monocarboxylates (SMCT2). Biochem J 392:655–664CrossRefGoogle Scholar
  9. 9.
    Gopal E, Umapathy NS, Martin PM, Ananth S, Gnana-Prakasam JP, Becker H, Wagner CA, Ganapathy V, Prasad PD (2007) Cloning and functional characterization of human SMCT2 (SLC5A12) and expression pattern of the transporter in kidney. Biochim Biophys Acta 1768:2690–2697CrossRefGoogle Scholar
  10. 10.
    Thangaraju M, Ananth S, Martin PM, Roon P, Smith SB, Sterneck E, Prasad PD, Ganapathy V (2006) c/ebpδ null mouse as a model for the double-knockout of slc5a8 and slc5a12 in kidney. J Biol Chem 281:26769–26773CrossRefGoogle Scholar
  11. 11.
    Enomoto A, Kimura H, Chairoungdua A, Shigeta Y, Jutabha P, Cha SH, Hosoyamada M, Takeda M, Sekine T, Igarashi T, Matsuo H, Kikuchi Y, Oda T, Ichida K, Hosoya T, Shimokata K, Niwa T, Kanai Y, Endou H (2002) Molecular identification of a renal urate anion exchanger that regulates blood urate levels. Nature 417:447–452CrossRefGoogle Scholar
  12. 12.
    Anzai N, Endou H (2007) Drug discovery for hyperuricemia. Expert Opin Drug Discov 2:1251–1261CrossRefGoogle Scholar
  13. 13.
    Ganapathy V, Gopal E, Miyauchi S, Prasad PD (2005) Biological functions of SLC5A8, a candidate tumour suppressor. Biochem Soc Trans 33:237–240CrossRefGoogle Scholar
  14. 14.
    Li H, Myeroff L, Smiraglia D, Romero MF, Pretlow TP, Kasturi L, Lutterbaugh J, Rerko RM, Casey G, Issa JP, Willis J, Willson JK, Plass C, Markowitz SD (2003) SLC5A8, a sodium transporter, is a tumor suppressor gene silenced by methylation in human colon aberrant crypt foci and cancers. Proc Natl Acad Sci USA 100:8412–8417CrossRefGoogle Scholar
  15. 15.
    Biber J (2001) Emerging roles of transporter-PDZ complexes in renal proximal tubular reabsorption. Pflugers Arch 443:3–5CrossRefGoogle Scholar
  16. 16.
    Levi M (2003) Role of PDZ domain-containing proteins and ERM proteins in regulation of renal function and dysfunction. J Am Soc Nephrol 14:1949–1951CrossRefGoogle Scholar
  17. 17.
    Moe OW (2003) Scaffolds: orchestrating proteins to achieve concerted function. Kidney Int 64:1916–1917CrossRefGoogle Scholar
  18. 18.
    Anzai N, Jutabha P, Kanai Y, Endou H (2005) Integrated physiology of proximal tubular organic anion transport. Curr Opin Nephrol Hypertens 14:472–479CrossRefGoogle Scholar
  19. 19.
    Fanning AS, Anderson JM (1999) Protein modules as organizers of membrane structure. Curr Opin Cell Biol 11:432–439CrossRefGoogle Scholar
  20. 20.
    Garner CC, Nash J, Huganir RL (2000) PDZ domains in synapse assembly and signalling. Trends Cell Biol 10:274–280CrossRefGoogle Scholar
  21. 21.
    Hung AY, Sheng M (2002) PDZ domains: structural modules for protein complex assembly. J Biol Chem 277:5699–5702CrossRefGoogle Scholar
  22. 22.
    Russel FGM, Masereeuw R, van Aubel RAMH (2002) Molecular aspects of renal anionic drug transport. Annu Rev Physiol 64:563–594CrossRefGoogle Scholar
  23. 23.
    Anzai N, Miyazaki H, Noshiro R, Khamdang S, Chairoungdua A, Shin HJ, Enomoto A, Sakamoto S, Hirata T, Tomita K, Kanai Y, Endou H (2004) The multivalent PDZ domain-containing protein PDZK1 regulates transport activity of renal urate-anion exchanger URAT1 via its C-terminal. J Biol Chem 279:45942–45950CrossRefGoogle Scholar
  24. 24.
    Noshiro R, Anzai N, Sakata T, Miyazaki H, Terada T, Shin HJ, He X, Miura D, Inui K, Kanai Y, Endou H (2006) The PDZ domain protein PDZK1 interacts with human peptide transporter PEPT2 and enhances its transport activity. Kidney Int 70:275–282CrossRefGoogle Scholar
  25. 25.
    Miyazaki H, Anzai N, Ekaratanawong S, Sakata T, Shin HJ, Jutabha P, Hirata T, He X, Nonoguchi H, Tomita K, Kanai Y, Endou H (2005) Modulation of renal apical organic anion transporter 4 (OAT4) function by two PDZ domain-containing proteins. J Am Soc Nephrol 16:3498–3506CrossRefGoogle Scholar
  26. 26.
    Kocher O, Comella N, Tognazzi K, Brown LF (1998) Identification and partial characterization of PDZK1: a novel protein containing PDZ interaction domains. Lab Investig 78:117–125Google Scholar
  27. 27.
    Weinman EJ, Steplock D, Wang Y, Shenolikar S (1995) Characterization of a protein cofactor that mediates protein kinase A regulation of the renal brush border membrane Na+–H+ exchanger. J Clin Investig 95:2143–2149CrossRefGoogle Scholar
  28. 28.
    Reczek D, Berryman M, Bretscher J (1997) Identification of EBP50: a PDZ-containing phosphoprotein that associates with members of the ezrin-radixin-moesin family. J Cell Biol 139:169–179CrossRefGoogle Scholar
  29. 29.
    Hall RA, Ostedgaard LS, Premont RT, Blitzer JT, Rahman N, Welsh MJ, Lefkowitz RJ (1998) A C-terminal motif found in the beta2-adrenergic receptor, P2Y1 receptor and cystic fibrosis transmembrane conductance regulator determines binding to the Na+/H+ exchanger regulatory factor family of PDZ proteins. Proc Natl Acad Sci USA 95:8496–8501CrossRefGoogle Scholar
  30. 30.
    Scott RO, Thelin WR, Milgram SL (2002) A novel PDZ protein regulates the activity of guanylyl cyclase C, the heat-stable enterotoxin receptor. J Biol Chem 277:22934–22941CrossRefGoogle Scholar
  31. 31.
    Songyang Z, Fanning AS, Fu C, Xu J, Marfatia SM, Chishti AH, Crompton A, Chan AC, Anderson JM, Cantley LC (1997) Recognition of unique carboxyl-terminal motifs by distinct PDZ domains. Science 275:73–77CrossRefGoogle Scholar
  32. 32.
    Kocher O, Comella N, Gilchrist A, Pal R, Tognazzi K, Brown LF, Knoll JH (1999) PDZK1, a novel PDZ domain-containing protein up-regulated in carcinomas and mapped to chromosome 1q21, interacts with cMOAT (MRP2), the multidrug resistance-associated protein. Lab Investig 79:1161–1170Google Scholar
  33. 33.
    Gisler SM, Pribanic S, Bacic D, Forrer P, Gantenbein A, Sabourin LA, Tsuji A, Zhao ZS, Manser E, Biber J, Murer H (2003) PDZK1: I. A major scaffolder in brush borders of proximal tubular cells. Kidney Int 64:1733–1745CrossRefGoogle Scholar
  34. 34.
    Anzai N, Kanai Y, Endou H (2007) New insights into renal transport of urate. Curr Opin Rheumatol 19:151–157CrossRefGoogle Scholar

Copyright information

© The Physiological Society of Japan and Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Pharmacology and ToxicologyKyorin University School of MedicineMitakaJapan
  2. 2.Department of PharmacologyChiba University Graduate School of MedicineChibaJapan
  3. 3.School of Chinese Materia MedicaTianjin University of Traditional Chinese MedicineTianjinChina
  4. 4.Department of Pharmaceutics, Faculty of Pharmaceutical SciencesToho UniversityFunabashiJapan

Personalised recommendations