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. 1998 May 15;101(10):2257–2267. doi: 10.1172/JCI2303

Defective aquaporin-2 trafficking in nephrogenic diabetes insipidus and correction by chemical chaperones.

B K Tamarappoo 1, A S Verkman 1
PMCID: PMC508814  PMID: 9593782

Abstract

Five single-point aquaporin-2 (AQP2) mutations that cause non-X-linked nephrogenic diabetes insipidus (NDI) were characterized to establish the cellular defect and to develop therapeutic strategies. In Xenopus oocytes expressing AQP2 cRNAs, single-channel water permeabilities of mutants L22V, T126M, and A147T were similar to that of wild-type AQP2, whereas R187C and C181W were nonfunctional. In [35S]methionine pulse-chase experiments in transiently transfected CHO cells, half-times for AQP2 degradation were approximately 4 h for wild-type AQP2 and L22V, and mildly decreased for T126M (2.7 h), C181W (2.4 h), R187C (2.0 h), and A147T (1.8 h). Immunofluorescence showed three distinct AQP2-staining patterns: plasma membrane and endosomal staining (wild-type, L22V), endoplasmic reticulum (ER) staining (T126M > A147T approximately R187C), or a mixed pattern of reticular and perinuclear vesicular staining. Immunoblot of fractionated vesicles confirmed primary ER localization of T126M, R187C, and A147T. To determine if the AQP2-trafficking defect is correctable, cells were incubated with the "chemical chaperone" glycerol for 48 h. Immunoblot showed that glycerol produced a nearly complete redistribution of AQP2 (T126M, A147T, and R187C) from ER to membrane/endosome fractions. Immunofluorescence confirmed the cellular redistribution. Redistribution of AQP2 mutants was also demonstrated in transfected MDCK cells, and using the chaperones TMAO and DMSO in place of glycerol in CHO cells. Water permeability measurements indicated that functional correction was achieved. These results indicate defective mammalian cell processing of mutant AQP2 water channels in NDI, and provide evidence for pharmacological correction of the processing defect by chemical chaperones.

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Selected References

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  1. Balch W. E., Dunphy W. G., Braell W. A., Rothman J. E. Reconstitution of the transport of protein between successive compartments of the Golgi measured by the coupled incorporation of N-acetylglucosamine. Cell. 1984 Dec;39(2 Pt 1):405–416. doi: 10.1016/0092-8674(84)90019-9. [DOI] [PubMed] [Google Scholar]
  2. Bergeron J. J., Brenner M. B., Thomas D. Y., Williams D. B. Calnexin: a membrane-bound chaperone of the endoplasmic reticulum. Trends Biochem Sci. 1994 Mar;19(3):124–128. doi: 10.1016/0968-0004(94)90205-4. [DOI] [PubMed] [Google Scholar]
  3. Bonifacino J. S., Lippincott-Schwartz J. Degradation of proteins within the endoplasmic reticulum. Curr Opin Cell Biol. 1991 Aug;3(4):592–600. doi: 10.1016/0955-0674(91)90028-w. [DOI] [PubMed] [Google Scholar]
  4. Brown C. R., Hong-Brown L. Q., Biwersi J., Verkman A. S., Welch W. J. Chemical chaperones correct the mutant phenotype of the delta F508 cystic fibrosis transmembrane conductance regulator protein. Cell Stress Chaperones. 1996 Jun;1(2):117–125. doi: 10.1379/1466-1268(1996)001<0117:ccctmp>2.3.co;2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown C. R., Hong-Brown L. Q., Welch W. J. Correcting temperature-sensitive protein folding defects. J Clin Invest. 1997 Mar 15;99(6):1432–1444. doi: 10.1172/JCI119302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown D. Membrane recycling and epithelial cell function. Am J Physiol. 1989 Jan;256(1 Pt 2):F1–12. doi: 10.1152/ajprenal.1989.256.1.F1. [DOI] [PubMed] [Google Scholar]
  7. Canfield M. C., Tamarappoo B. K., Moses A. M., Verkman A. S., Holtzman E. J. Identification and characterization of aquaporin-2 water channel mutations causing nephrogenic diabetes insipidus with partial vasopressin response. Hum Mol Genet. 1997 Oct;6(11):1865–1871. doi: 10.1093/hmg/6.11.1865. [DOI] [PubMed] [Google Scholar]
  8. Deen P. M., Croes H., van Aubel R. A., Ginsel L. A., van Os C. H. Water channels encoded by mutant aquaporin-2 genes in nephrogenic diabetes insipidus are impaired in their cellular routing. J Clin Invest. 1995 May;95(5):2291–2296. doi: 10.1172/JCI117920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deen P. M., Verdijk M. A., Knoers N. V., Wieringa B., Monnens L. A., van Os C. H., van Oost B. A. Requirement of human renal water channel aquaporin-2 for vasopressin-dependent concentration of urine. Science. 1994 Apr 1;264(5155):92–95. doi: 10.1126/science.8140421. [DOI] [PubMed] [Google Scholar]
  10. Denning G. M., Anderson M. P., Amara J. F., Marshall J., Smith A. E., Welsh M. J. Processing of mutant cystic fibrosis transmembrane conductance regulator is temperature-sensitive. Nature. 1992 Aug 27;358(6389):761–764. doi: 10.1038/358761a0. [DOI] [PubMed] [Google Scholar]
  11. Edington B. V., Whelan S. A., Hightower L. E. Inhibition of heat shock (stress) protein induction by deuterium oxide and glycerol: additional support for the abnormal protein hypothesis of induction. J Cell Physiol. 1989 May;139(2):219–228. doi: 10.1002/jcp.1041390202. [DOI] [PubMed] [Google Scholar]
  12. Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farinas J., Simanek V., Verkman A. S. Cell volume measured by total internal reflection microfluorimetry: application to water and solute transport in cells transfected with water channel homologs. Biophys J. 1995 Apr;68(4):1613–1620. doi: 10.1016/S0006-3495(95)80335-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fujiwara T. M., Morgan K., Bichet D. G. Molecular biology of diabetes insipidus. Annu Rev Med. 1995;46:331–343. doi: 10.1146/annurev.med.46.1.331. [DOI] [PubMed] [Google Scholar]
  15. Fushimi K., Uchida S., Hara Y., Hirata Y., Marumo F., Sasaki S. Cloning and expression of apical membrane water channel of rat kidney collecting tubule. Nature. 1993 Feb 11;361(6412):549–552. doi: 10.1038/361549a0. [DOI] [PubMed] [Google Scholar]
  16. Gekko K., Koga S. Increased thermal stability of collagen in the presence of sugars and polyols. J Biochem. 1983 Jul;94(1):199–205. doi: 10.1093/oxfordjournals.jbchem.a134330. [DOI] [PubMed] [Google Scholar]
  17. Gekko K., Timasheff S. N. Thermodynamic and kinetic examination of protein stabilization by glycerol. Biochemistry. 1981 Aug 4;20(16):4677–4686. doi: 10.1021/bi00519a024. [DOI] [PubMed] [Google Scholar]
  18. Jung J. S., Preston G. M., Smith B. L., Guggino W. B., Agre P. Molecular structure of the water channel through aquaporin CHIP. The hourglass model. J Biol Chem. 1994 May 20;269(20):14648–14654. [PubMed] [Google Scholar]
  19. Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
  20. Langley J. M., Balfe J. W., Selander T., Ray P. N., Clarke J. T. Autosomal recessive inheritance of vasopressin-resistant diabetes insipidus. Am J Med Genet. 1991 Jan;38(1):90–94. doi: 10.1002/ajmg.1320380120. [DOI] [PubMed] [Google Scholar]
  21. Marsh E. W., Leopold P. L., Jones N. L., Maxfield F. R. Oligomerized transferrin receptors are selectively retained by a lumenal sorting signal in a long-lived endocytic recycling compartment. J Cell Biol. 1995 Jun;129(6):1509–1522. doi: 10.1083/jcb.129.6.1509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Melnick J., Aviel S., Argon Y. The endoplasmic reticulum stress protein GRP94, in addition to BiP, associates with unassembled immunoglobulin chains. J Biol Chem. 1992 Oct 25;267(30):21303–21306. [PubMed] [Google Scholar]
  23. Mulders S. M., Knoers N. V., Van Lieburg A. F., Monnens L. A., Leumann E., Wühl E., Schober E., Rijss J. P., Van Os C. H., Deen P. M. New mutations in the AQP2 gene in nephrogenic diabetes insipidus resulting in functional but misrouted water channels. J Am Soc Nephrol. 1997 Feb;8(2):242–248. doi: 10.1681/ASN.V82242. [DOI] [PubMed] [Google Scholar]
  24. Nielsen S., Chou C. L., Marples D., Christensen E. I., Kishore B. K., Knepper M. A. Vasopressin increases water permeability of kidney collecting duct by inducing translocation of aquaporin-CD water channels to plasma membrane. Proc Natl Acad Sci U S A. 1995 Feb 14;92(4):1013–1017. doi: 10.1073/pnas.92.4.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pan Y., Metzenberg A., Das S., Jing B., Gitschier J. Mutations in the V2 vasopressin receptor gene are associated with X-linked nephrogenic diabetes insipidus. Nat Genet. 1992 Oct;2(2):103–106. doi: 10.1038/ng1092-103. [DOI] [PubMed] [Google Scholar]
  26. Pind S., Riordan J. R., Williams D. B. Participation of the endoplasmic reticulum chaperone calnexin (p88, IP90) in the biogenesis of the cystic fibrosis transmembrane conductance regulator. J Biol Chem. 1994 Apr 29;269(17):12784–12788. [PubMed] [Google Scholar]
  27. Qu B. H., Strickland E. H., Thomas P. J. Localization and suppression of a kinetic defect in cystic fibrosis transmembrane conductance regulator folding. J Biol Chem. 1997 Jun 20;272(25):15739–15744. doi: 10.1074/jbc.272.25.15739. [DOI] [PubMed] [Google Scholar]
  28. Sabolić I., Wuarin F., Shi L. B., Verkman A. S., Ausiello D. A., Gluck S., Brown D. Apical endosomes isolated from kidney collecting duct principal cells lack subunits of the proton pumping ATPase. J Cell Biol. 1992 Oct;119(1):111–122. doi: 10.1083/jcb.119.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sato S., Ward C. L., Krouse M. E., Wine J. J., Kopito R. R. Glycerol reverses the misfolding phenotype of the most common cystic fibrosis mutation. J Biol Chem. 1996 Jan 12;271(2):635–638. doi: 10.1074/jbc.271.2.635. [DOI] [PubMed] [Google Scholar]
  30. Storrie B., Madden E. A. Isolation of subcellular organelles. Methods Enzymol. 1990;182:203–225. doi: 10.1016/0076-6879(90)82018-w. [DOI] [PubMed] [Google Scholar]
  31. Tatzelt J., Prusiner S. B., Welch W. J. Chemical chaperones interfere with the formation of scrapie prion protein. EMBO J. 1996 Dec 2;15(23):6363–6373. [PMC free article] [PubMed] [Google Scholar]
  32. Wall D. A., Patel S. Isolation of plasma membrane complexes from Xenopus oocytes. J Membr Biol. 1989 Feb;107(2):189–201. doi: 10.1007/BF01871724. [DOI] [PubMed] [Google Scholar]
  33. Ward C. L., Omura S., Kopito R. R. Degradation of CFTR by the ubiquitin-proteasome pathway. Cell. 1995 Oct 6;83(1):121–127. doi: 10.1016/0092-8674(95)90240-6. [DOI] [PubMed] [Google Scholar]
  34. Welch W. J., Brown C. R. Influence of molecular and chemical chaperones on protein folding. Cell Stress Chaperones. 1996 Jun;1(2):109–115. doi: 10.1379/1466-1268(1996)001<0109:iomacc>2.3.co;2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Welsh M. J., Smith A. E. Molecular mechanisms of CFTR chloride channel dysfunction in cystic fibrosis. Cell. 1993 Jul 2;73(7):1251–1254. doi: 10.1016/0092-8674(93)90353-r. [DOI] [PubMed] [Google Scholar]
  36. Yang Y., Janich S., Cohn J. A., Wilson J. M. The common variant of cystic fibrosis transmembrane conductance regulator is recognized by hsp70 and degraded in a pre-Golgi nonlysosomal compartment. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9480–9484. doi: 10.1073/pnas.90.20.9480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Zhang R., Skach W., Hasegawa H., van Hoek A. N., Verkman A. S. Cloning, functional analysis and cell localization of a kidney proximal tubule water transporter homologous to CHIP28. J Cell Biol. 1993 Jan;120(2):359–369. doi: 10.1083/jcb.120.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. van Lieburg A. F., Verdijk M. A., Knoers V. V., van Essen A. J., Proesmans W., Mallmann R., Monnens L. A., van Oost B. A., van Os C. H., Deen P. M. Patients with autosomal nephrogenic diabetes insipidus homozygous for mutations in the aquaporin 2 water-channel gene. Am J Hum Genet. 1994 Oct;55(4):648–652. [PMC free article] [PubMed] [Google Scholar]

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