Abstract
Two forms of intercalated cells are present in kidney collecting tubules, the alpha cell has apical endocytosis, apical H+-ATPase and basolateral band 3, while beta cells have reversed polarity of these proteins and no apical endocytosis. When a beta cell line was seeded at high density, it changed into the alpha form. We previously showed that a partially purified 230 kD extracellular matrix protein of high density cells was able to retarget band 3 from apical to basolateral domains and stimulated apical endocytosis in vitro (Van Adelsberg, J., J.C. Edwards, J. Takito, B. Kiss, and Q. Al-Awqati. 1994. Cell. 76:1053-1061). We now purify this protein, which was named hensin, to near homogeneity and find that it belongs to the macrophage scavenger receptor cysteine rich (SRCR) family. An antibody, generated against a fusion protein made from a partial cDNA recognized a 230-kD protein in rabbit kidney and in the intercalated cell line. In vitro, the hensin antibody inhibited expression of apical endocytosis. Hensin was secreted in a polarized manner and bound to the basolateral membrane and extracellular matrix. Immunohistochemistry of the kidney showed that it was expressed only in collecting tubules. Double immunofluorescence with hensin and peanut lectin, H+-ATPase, or band 3 showed many patterns; most alpha-cells had hensin staining while 50% of beta-cells did not. These results suggest that hensin may also be involved in the polarity reversal of intercalated cells in vivo.
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- Al-Awqati Q. Plasticity in epithelial polarity of renal intercalated cells: targeting of the H(+)-ATPase and band 3. Am J Physiol. 1996 Jun;270(6 Pt 1):C1571–C1580. doi: 10.1152/ajpcell.1996.270.6.C1571. [DOI] [PubMed] [Google Scholar]
- Alper S. L., Natale J., Gluck S., Lodish H. F., Brown D. Subtypes of intercalated cells in rat kidney collecting duct defined by antibodies against erythroid band 3 and renal vacuolar H+-ATPase. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5429–5433. doi: 10.1073/pnas.86.14.5429. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bastani B., Purcell H., Hemken P., Trigg D., Gluck S. Expression and distribution of renal vacuolar proton-translocating adenosine triphosphatase in response to chronic acid and alkali loads in the rat. J Clin Invest. 1991 Jul;88(1):126–136. doi: 10.1172/JCI115268. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown D., Hirsch S., Gluck S. An H+-ATPase in opposite plasma membrane domains in kidney epithelial cell subpopulations. Nature. 1988 Feb 18;331(6157):622–624. doi: 10.1038/331622a0. [DOI] [PubMed] [Google Scholar]
- Chicheportiche Y., Vassalli P. Cloning and expression of a mouse macrophage cDNA coding for a membrane glycoprotein of the scavenger receptor cysteine-rich domain family. J Biol Chem. 1994 Feb 25;269(8):5512–5517. [PubMed] [Google Scholar]
- Drenckhahn D., Schlüter K., Allen D. P., Bennett V. Colocalization of band 3 with ankyrin and spectrin at the basal membrane of intercalated cells in the rat kidney. Science. 1985 Dec 13;230(4731):1287–1289. doi: 10.1126/science.2933809. [DOI] [PubMed] [Google Scholar]
- Edwards J. C., van Adelsberg J., Rater M., Herzlinger D., Lebowitz J., al-Awqati Q. Conditional immortalization of bicarbonate-secreting intercalated cells from rabbit. Am J Physiol. 1992 Aug;263(2 Pt 1):C521–C529. doi: 10.1152/ajpcell.1992.263.2.C521. [DOI] [PubMed] [Google Scholar]
- Elomaa O., Kangas M., Sahlberg C., Tuukkanen J., Sormunen R., Liakka A., Thesleff I., Kraal G., Tryggvason K. Cloning of a novel bacteria-binding receptor structurally related to scavenger receptors and expressed in a subset of macrophages. Cell. 1995 Feb 24;80(4):603–609. doi: 10.1016/0092-8674(95)90514-6. [DOI] [PubMed] [Google Scholar]
- Fejes-Tóth G., Náray-Fejes-Tóth A. Differentiation of renal beta-intercalated cells to alpha-intercalated and principal cells in culture. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5487–5491. doi: 10.1073/pnas.89.12.5487. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Freeman M., Ashkenas J., Rees D. J., Kingsley D. M., Copeland N. G., Jenkins N. A., Krieger M. An ancient, highly conserved family of cysteine-rich protein domains revealed by cloning type I and type II murine macrophage scavenger receptors. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8810–8814. doi: 10.1073/pnas.87.22.8810. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jennings M. L., Anderson M. P., Monaghan R. Monoclonal antibodies against human erythrocyte band 3 protein. Localization of proteolytic cleavage sites and stilbenedisulfonate-binding lysine residues. J Biol Chem. 1986 Jul 5;261(19):9002–9010. [PubMed] [Google Scholar]
- Kemperman H., Wijnands Y., Wesseling J., Niessen C. M., Sonnenberg A., Roos E. The mucin epiglycanin on TA3/Ha carcinoma cells prevents alpha 6 beta 4-mediated adhesion to laminin and kalinin and E-cadherin-mediated cell-cell interaction. J Cell Biol. 1994 Dec;127(6 Pt 2):2071–2080. doi: 10.1083/jcb.127.6.2071. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LeHir M., Kaissling B., Koeppen B. M., Wade J. B. Binding of peanut lectin to specific epithelial cell types in kidney. Am J Physiol. 1982 Jan;242(1):C117–C120. doi: 10.1152/ajpcell.1982.242.1.C117. [DOI] [PubMed] [Google Scholar]
- Minuth W. W., Gilbert P., Rudolph U., Spielman W. S. Successive histochemical differentiation steps during postnatal development of the collecting duct in rabbit kidney. Histochemistry. 1989;93(1):19–25. doi: 10.1007/BF00266842. [DOI] [PubMed] [Google Scholar]
- Resnick D., Pearson A., Krieger M. The SRCR superfamily: a family reminiscent of the Ig superfamily. Trends Biochem Sci. 1994 Jan;19(1):5–8. doi: 10.1016/0968-0004(94)90165-1. [DOI] [PubMed] [Google Scholar]
- Romano V., Hatzfeld M., Magin T. M., Zimbelmann R., Franke W. W., Maier G., Ponstingl H. Cytokeratin expression in simple epithelia. I. Identification of mRNA coding for human cytokeratin no. 18 by a cDNA clone. Differentiation. 1986;30(3):244–253. doi: 10.1111/j.1432-0436.1986.tb00787.x. [DOI] [PubMed] [Google Scholar]
- Schuster V. L. Function and regulation of collecting duct intercalated cells. Annu Rev Physiol. 1993;55:267–288. doi: 10.1146/annurev.ph.55.030193.001411. [DOI] [PubMed] [Google Scholar]
- Schwartz G. J., Barasch J., Al-Awqati Q. Plasticity of functional epithelial polarity. 1985 Nov 28-Dec 4Nature. 318(6044):368–371. doi: 10.1038/318368a0. [DOI] [PubMed] [Google Scholar]
- Steinmetz P. R. Cellular organization of urinary acidification. Am J Physiol. 1986 Aug;251(2 Pt 2):F173–F187. doi: 10.1152/ajprenal.1986.251.2.F173. [DOI] [PubMed] [Google Scholar]
- Ullrich A., Sures I., D'Egidio M., Jallal B., Powell T. J., Herbst R., Dreps A., Azam M., Rubinstein M., Natoli C. The secreted tumor-associated antigen 90K is a potent immune stimulator. J Biol Chem. 1994 Jul 15;269(28):18401–18407. [PubMed] [Google Scholar]
- Van Adelsberg J., Edwards J. C., Herzlinger D., Cannon C., Rater M., al-Awqati Q. Isolation and culture of HCO3- -secreting intercalated cells. Am J Physiol. 1989 May;256(5 Pt 1):C1004–C1011. doi: 10.1152/ajpcell.1989.256.5.C1004. [DOI] [PubMed] [Google Scholar]
- Verlander J. W., Madsen K. M., Cannon J. K., Tisher C. C. Activation of acid-secreting intercalated cells in rabbit collecting duct with ammonium chloride loading. Am J Physiol. 1994 Apr;266(4 Pt 2):F633–F645. doi: 10.1152/ajprenal.1994.266.4.F633. [DOI] [PubMed] [Google Scholar]
- Whitney G. S., Starling G. C., Bowen M. A., Modrell B., Siadak A. W., Aruffo A. The membrane-proximal scavenger receptor cysteine-rich domain of CD6 contains the activated leukocyte cell adhesion molecule binding site. J Biol Chem. 1995 Aug 4;270(31):18187–18190. doi: 10.1074/jbc.270.31.18187. [DOI] [PubMed] [Google Scholar]
- Wijngaard P. L., Metzelaar M. J., MacHugh N. D., Morrison W. I., Clevers H. C. Molecular characterization of the WC1 antigen expressed specifically on bovine CD4-CD8- gamma delta T lymphocytes. J Immunol. 1992 Nov 15;149(10):3273–3277. [PubMed] [Google Scholar]
- Yurko M. A., Gluck S. Production and characterization of a monoclonal antibody to vacuolar H+ATPase of renal epithelia. J Biol Chem. 1987 Nov 15;262(32):15770–15779. [PubMed] [Google Scholar]
- van Adelsberg J. S., Edwards J. C., al-Awqati Q. The apical Cl/HCO3 exchanger of beta intercalated cells. J Biol Chem. 1993 May 25;268(15):11283–11289. [PubMed] [Google Scholar]
- van Adelsberg J., Edwards J. C., Takito J., Kiss B., al-Awqati Q. An induced extracellular matrix protein reverses the polarity of band 3 in intercalated epithelial cells. Cell. 1994 Mar 25;76(6):1053–1061. doi: 10.1016/0092-8674(94)90382-4. [DOI] [PubMed] [Google Scholar]