Synthesis, Maturation, and Trafficking of Human Na+-Dicarboxylate Cotransporter NaDC1 Requires the Chaperone Activity of Cyclophilin B*

Renal excretion of citrate, an inhibitor of calcium stone formation, is controlled mainly by reabsorption via the apical Na+-dicarboxylate cotransporter NaDC1 (SLC13A2) in the proximal tubule. Recently, it has been shown that the protein phosphatase calcineurin inhibitors cyclosporin A (CsA) and FK-506 induce hypocitraturia, a risk factor for nephrolithiasis in kidney transplant patients, but apparently through urine acidification. This suggests that these agents up-regulate NaDC1 activity. Using the Xenopus lævis oocyte and HEK293 cell expression systems, we examined first the effect of both anti-calcineurins on NaDC1 activity and expression. While FK-506 had no effect, CsA reduced NaDC1-mediated citrate transport by lowering heterologous carrier expression (as well as endogenous carrier expression in HEK293 cells), indicating that calcineurin is not involved. Given that CsA also binds specifically to cyclophilins, we determined next whether such proteins could account for the observed changes by examining the effect of selected cyclophilin wild types and mutants on NaDC1 activity and cyclophilin-specific siRNA. Interestingly, our data show that the cyclophilin isoform B is likely responsible for down-regulation of carrier expression by CsA and that it does so via its chaperone activity on NaDC1 (by direct interaction) rather than its rotamase activity. We have thus identified for the first time a regulatory partner for NaDC1, and have gained novel mechanistic insight into the effect of CsA on renal citrate transport and kidney stone disease, as well as into the regulation of membrane transporters in general.

[1]  J. Luban,et al.  Cyclophilin B Interacts with Sodium-Potassium ATPase and Is Required for Pump Activity in Proximal Tubule Cells of the Kidney , 2010, PloS one.

[2]  N. Zheleznova,et al.  Calcineurin Interacts with PERK and Dephosphorylates Calnexin to Relieve ER Stress in Mammals and Frogs , 2010, PloS one.

[3]  G. Salido,et al.  SERCA2b Activity Is Regulated by Cyclophilins in Human Platelets , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[4]  P. Beaune,et al.  Cyclosporine‐Induced Endoplasmic Reticulum Stress Triggers Tubular Phenotypic Changes and Death , 2008, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[5]  U. Wissenbach,et al.  The Human TRPV6 Channel Protein Is Associated with Cyclophilin B in Human Placenta* , 2008, Journal of Biological Chemistry.

[6]  M. Gottesman,et al.  Modulation of Na+-Ca2+ Exchanger Expression by Immunosuppressive Drugs Is Isoform-Specific , 2008, Molecular Pharmacology.

[7]  R. Wek,et al.  Translational control and the unfolded protein response. , 2007, Antioxidants & redox signaling.

[8]  T. Hunt,et al.  Calcineurin is required to release Xenopus egg extracts from meiotic M phase. , 2007, Nature.

[9]  A. Cheung,et al.  Generation and characterization of sodium-dicarboxylate cotransporter-deficient mice. , 2007, Kidney international.

[10]  T. Kitamura,et al.  Associations between renal sodium‐citrate cotransporter (hNaDC‐1) gene polymorphism and urinary citrate excretion in recurrent renal calcium stone formers and normal controls , 2007, International journal of urology : official journal of the Japanese Urological Association.

[11]  S. Sengul,et al.  Renal tubular acidosis after kidney transplantation--incidence, risk factors and clinical implications. , 2007, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[12]  Junying Yuan,et al.  Structure-activity relationship studies of salubrinal lead to its active biotinylated derivative. , 2005, Bioorganic & medicinal chemistry letters.

[13]  M. Bukrinsky,et al.  Regulation of CD147 Cell Surface Expression , 2005, Journal of Biological Chemistry.

[14]  B. Hoppe,et al.  Hypocitraturia as a risk factor for nephrocalcinosis after kidney transplantation , 2005, Pediatric Nephrology.

[15]  Jeong-Sun Seo,et al.  Transgenic mice overexpressing cyclophilin A are resistant to cyclosporin A-induced nephrotoxicity via peptidyl-prolyl cis-trans isomerase activity. , 2004, Biochemical and biophysical research communications.

[16]  J. Tejada,et al.  Continuous Recording of Blood Pressure in Cerebrovascular Disease: When Should It Be Done? , 2003, Cerebrovascular Diseases.

[17]  Andrzej Galat,et al.  Peptidylprolyl cis/trans isomerases (immunophilins): biological diversity--targets--functions. , 2003, Current topics in medicinal chemistry.

[18]  L. Hamm,et al.  Pathophysiology of hypocitraturic nephrolithiasis. , 2002, Endocrinology and metabolism clinics of North America.

[19]  C. Clevenger,et al.  The intranuclear prolactin/cyclophilin B complex as a transcriptional inducer , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[20]  H. Rahamimoff,et al.  Transport Activity and Surface Expression of the Na+-Ca2+ Exchanger NCX1 Are Inhibited by the Immunosuppressive Agent Cyclosporin A and by the Nonimmunosuppressive Agent PSC833* , 2002, The Journal of Biological Chemistry.

[21]  H. Sarkar,et al.  Cyclosporin A Inhibits Creatine Uptake by Altering Surface Expression of the Creatine Transporter* , 2000, The Journal of Biological Chemistry.

[22]  Y. Uezono,et al.  Post-translational reduction of cell surface expression of insulin receptors by cyclosporin A, FK506 and rapamycin in bovine adrenal chromaffin cells , 2000, Neuroscience Letters.

[23]  S. Wehrli,et al.  Chronic metabolic acidosis increases NaDC-1 mRNA and protein abundance in rat kidney. , 2000, Kidney international.

[24]  Florian Lang,et al.  The Use of Xenopus laevis Oocytes for the Functional Characterization of Heterologously Expressed Membrane Proteins , 2000, Cellular Physiology and Biochemistry.

[25]  S. Heinemann,et al.  Cyclosporin A selectively reduces the functional expression of Kir2.1 potassium channels in Xenopus oocytes , 1998, FEBS letters.

[26]  S. Helekar,et al.  Peptidyl prolyl cis-trans isomerase activity of cyclophilin A in functional homo-oligomeric receptor expression. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[27]  C. Zuker,et al.  The cyclophilin homolog NinaA functions as a chaperone, forming a stable complex in vivo with its protein target rhodopsin. , 1994, The EMBO journal.

[28]  C. Walsh,et al.  Cyclophilin B trafficking through the secretory pathway is altered by binding of cyclosporin A. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  C. Zuker,et al.  The cyclophilin homolog ninaA is required in the secretory pathway , 1991, Cell.

[30]  C. Walsh,et al.  Human cyclophilin B: a second cyclophilin gene encodes a peptidyl-prolyl isomerase with a signal sequence. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[31]  L. Hamm.  Renal handling of citrate. , 1990, Kidney international.

[32]  L. Shaw,et al.  Canadian Consensus Meeting on cyclosporine monitoring: report of the consensus panel. , 1990, Clinical chemistry.

[33]  Donker Aj,et al.  PROSPECTIVE SERIAL RENAL-FUNCTION STUDIES IN PATIENTS WITH NONRENAL DISEASE TREATED WITH CYCLOSPORINE-A , 1988 .

[34]  M. Okuhara,et al.  FK-506, a novel immunosuppressant isolated from a Streptomyces. I. Fermentation, isolation, and physico-chemical and biological characteristics. , 1987, The Journal of antibiotics.

[35]  L. Smith,et al.  Transport of citrate across renal brush border membrane: effects of dietary acid and alkali loading. , 1985, The American journal of physiology.

[36]  P. Beaune,et al.  Cyclosporine triggers endoplasmic reticulum stress in endothelial cells: a role for endothelial phenotypic changes and death. , 2009, American journal of physiology. Renal physiology.

[37]  H. Sarkar,et al.  Inhibition of taurine transport by cyclosporin A is due to altered surface abundance of the taurine transporter and is reversible. , 2009, Advances in experimental medicine and biology.

[38]  A. Fujimura,et al.  Cyclosporin A produces distal renal tubular acidosis by blocking peptidyl prolyl cis-trans isomerase activity of cyclophilin. , 2005, American journal of physiology. Renal physiology.

[39]  H. Gunshin,et al.  Expression cloning using Xenopus laevis oocytes. , 1998, Methods in enzymology.

[40]  P. Heering,et al.  Tubular dysfunction following kidney transplantation. , 1996, Nephron.