Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Oct;87(19):7532–7535. doi: 10.1073/pnas.87.19.7532

Proximal tubular epithelial cells possess a novel 42-kilodalton guanine nucleotide-binding regulatory protein.

J Zhou 1, C Sims 1, C H Chang 1, L Berti-Mattera 1, U Hopfer 1, J Douglas 1
PMCID: PMC54781  PMID: 2120702

Abstract

The proximal tubule of the kidney represents an important location where adenylate cyclase regulates salt and water transport; yet a detailed characterization of the distribution and classification of guanine nucleotide-binding protein (G protein) and adenylate cyclase is lacking. We used purified brush border (20-fold) and basolateral membranes (14-fold) to characterize parathyroid hormone- and G protein-regulated adenylate cyclase and G-protein distribution. Adenylate cyclase was predominantly localized to basolateral membranes, while the 46-kDa alpha subunit of the stimulatory G protein (Gs) was 2-fold higher in brush border membranes than in basolateral membranes. The alpha subunit of the inhibitory G protein (Gi; 41 kDa) was equally distributed on immunoblotting but was 2-fold higher in brush border membranes than in basolateral membranes on radiolabeling with pertussis toxin. A 42-kDa cholera toxin substrate that cross-reacted with antisera to the common alpha subunit of G proteins and to Gs on immunoblotting and that was not immunoprecipitated with two Gi antisera was the most abundant alpha subunit and comprised approximately 1% of the total membrane proteins. These observations suggest that G proteins are important regulators of proximal tubular transport independent of adenylate cyclase.

Full text

PDF
7532

Images in this article

7534Image
on p.7534

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berger S. J., Sacktor B. Isolation and biochemical characterization of brush borders from rabbit kidney. J Cell Biol. 1970 Dec;47(3):637–645. doi: 10.1083/jcb.47.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bokoch G. M., Katada T., Northup J. K., Hewlett E. L., Gilman A. G. Identification of the predominant substrate for ADP-ribosylation by islet activating protein. J Biol Chem. 1983 Feb 25;258(4):2072–2075. [PubMed] [Google Scholar]
  3. Burnatowska-Hledin M. A., Spielman W. S. Vasopressin V1 receptors on the principal cells of the rabbit cortical collecting tubule. Stimulation of cytosolic free calcium and inositol phosphate production via coupling to a pertussis toxin substrate. J Clin Invest. 1989 Jan;83(1):84–89. doi: 10.1172/JCI113888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carlson K. E., Brass L. F., Manning D. R. Thrombin and phorbol esters cause the selective phosphorylation of a G protein other than Gi in human platelets. J Biol Chem. 1989 Aug 5;264(22):13298–13305. [PubMed] [Google Scholar]
  5. Domínguez P., Barros F., Lazo P. S. The activation of adenylate cyclase from small intestinal epithelium by cholera toxin. Eur J Biochem. 1985 Feb 1;146(3):533–538. doi: 10.1111/j.1432-1033.1985.tb08684.x. [DOI] [PubMed] [Google Scholar]
  6. Domínguez P., Velasco G., Barros F., Lazo P. S. Intestinal brush border membranes contain regulatory subunits of adenylyl cyclase. Proc Natl Acad Sci U S A. 1987 Oct;84(20):6965–6969. doi: 10.1073/pnas.84.20.6965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Lazo P. S., Barros F., Domínguez P., Rivaya A., Velasco G. Adenylate cyclase from rabbit small intestine: activation by cholera toxin and interaction with calcium. Arch Biochem Biophys. 1985 Jun;239(2):587–594. doi: 10.1016/0003-9861(85)90728-3. [DOI] [PubMed] [Google Scholar]
  11. Light D. B., Ausiello D. A., Stanton B. A. Guanine nucleotide-binding protein, alpha i-3, directly activates a cation channel in rat renal inner medullary collecting duct cells. J Clin Invest. 1989 Jul;84(1):352–356. doi: 10.1172/JCI114162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Liu F. Y., Cogan M. G. Angiotensin II stimulates early proximal bicarbonate absorption in the rat by decreasing cyclic adenosine monophosphate. J Clin Invest. 1989 Jul;84(1):83–91. doi: 10.1172/JCI114174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Molina y Vedia L. M., Reep B. R., Lapetina E. G. Platelet cytosolic 44-kDa protein is a substrate of cholera toxin-induced ADP-ribosylation and is not recognized by antisera against the alpha subunit of the stimulatory guanine nucleotide-binding regulatory protein. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5899–5902. doi: 10.1073/pnas.85.16.5899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mumby S. M., Kahn R. A., Manning D. R., Gilman A. G. Antisera of designed specificity for subunits of guanine nucleotide-binding regulatory proteins. Proc Natl Acad Sci U S A. 1986 Jan;83(2):265–269. doi: 10.1073/pnas.83.2.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Murer H., Ammann E., Biber J., Hopfer U. The surface membrane of the small intestinal epithelial cell. I. Localization of adenyl cyclase. Biochim Biophys Acta. 1976 May 21;433(3):509–519. doi: 10.1016/0005-2736(76)90277-7. [DOI] [PubMed] [Google Scholar]
  16. Portilla D., Morrissey J., Morrison A. R. Bradykinin-activated membrane-associated phospholipase C in Madin-Darby canine kidney cells. J Clin Invest. 1988 Jun;81(6):1896–1902. doi: 10.1172/JCI113536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Scalera V., Huang Y. K., Hildmann B., Murer H. A simple isolation method for basal-lateral plasma membranes from rat kidney cortex. Membr Biochem. 1981;4(1):49–61. doi: 10.3109/09687688109065422. [DOI] [PubMed] [Google Scholar]
  18. Skorecki K. L., Verkman A. S., Ausiello D. A. Vasopressin receptor-adenylate cyclase interactions. Studies in an intact cultured renal epithelial cell line (LLC-PK1). Miner Electrolyte Metab. 1986;12(1):64–70. [PubMed] [Google Scholar]
  19. Szasz G. A kinetic photometric method for serum leucine aminopeptidase. Am J Clin Pathol. 1967 May;47(5):607–613. doi: 10.1093/ajcp/47.5.607. [DOI] [PubMed] [Google Scholar]
  20. Woolkalis M. J., Nakada M. T., Manning D. R. Alterations in components of adenylate cyclase associated with transformation of chicken embryo fibroblasts by Rous sarcoma virus. J Biol Chem. 1986 Mar 5;261(7):3408–3413. [PubMed] [Google Scholar]
  21. Yatani A., Mattera R., Codina J., Graf R., Okabe K., Padrell E., Iyengar R., Brown A. M., Birnbaumer L. The G protein-gated atrial K+ channel is stimulated by three distinct Gi alpha-subunits. Nature. 1988 Dec 15;336(6200):680–682. doi: 10.1038/336680a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES