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. 1982 Oct 1;95(1):8–19. doi: 10.1083/jcb.95.1.8

Plasma membrane of the rat parotid gland: preparation and partial characterization of a fraction containing the secretory surface

PMCID: PMC2112380  PMID: 6128347

Abstract

A plasma membrane fraction from the rat parotid gland has been prepared by a procedure which selectively enriches for large membrane sheets. This fraction appears to have preserved several ultrastructural features of the acinar cell surface observed in situ. Regions of membrane resembling the acinar luminal border appear as compartments containing microvillar invaginations, bounded by elements of the junctional complex, and from which basolateral membranes extend beyond the junctional complex either to contact other apical compartments or to terminate as free ends. Several additional morphological features of the apical compartments suggest that they are primarily derived from the surface of acinar cells, rather than from the minority of other salivary gland cell types. Enzymatic activities characteristically associated with other cellular organelles are found at only low levels in the plasma membrane fraction. The fraction is highly enriched in two enzyme activities--K+ -dependent p-nitrophenyl phosphatase (K+ -NPPase, shown to be Na+/K+ adenosine triphosphatase; 20-fold) and gamma- glutamyl transpeptidase (GGTPase; 26-fold)--both known to mark plasma membranes in other tissues. These activities exhibit different patterns of recovery during fractionation, suggesting their distinct distributions among parotid cellular membranes. Secretion granule membranes also exhibit GGTPase, but no detectable K+ -NPPase. Since Na+/K+ adenosine triphosphatase and GGTPase, respectively, mark the basolateral and apical cellular surfaces in other epithelia, we hypothesize that these two enzymes mark distinct domains in the parotid plasmalemma, and that GGTPase, as the putative apical marker, may signify a compositional overlap between the two types of membranes which fuse during exocytosis.

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Selected References

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  1. Adelman M. R., Blobel G., Sabatini D. D. An improved cell fractionation procedure for the preparation of rat liver membrane-bound ribosomes. J Cell Biol. 1973 Jan;56(1):191–205. doi: 10.1083/jcb.56.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amsterdam A., Schramm M., Ohad I., Salomon Y., Selinger Z. Concomitant synthesis of membrane protein and exportable protein of the secretory granule in rat parotid gland. J Cell Biol. 1971 Jul;50(1):187–200. doi: 10.1083/jcb.50.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  4. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  5. Bader H., Sen A. K. (K+)-dependent acyl phosphatase as part of the (na+ + K+)-dependent ATPase of cell membranes. Biochim Biophys Acta. 1966 Apr 12;118(1):116–123. doi: 10.1016/s0926-6593(66)80150-9. [DOI] [PubMed] [Google Scholar]
  6. Castle A. M., Castle J. D. The purification and partial characterization of phospholipase A2, a secretory protein of rabbit parotid gland. Biochim Biophys Acta. 1981 Nov 23;666(2):259–274. doi: 10.1016/0005-2760(81)90116-8. [DOI] [PubMed] [Google Scholar]
  7. Castle J. D., Jamieson J. D., Palade G. E. Secretion granules of the rabbit parotid gland. Isolation, subfractionation, and characterization of the membrane and content subfractions. J Cell Biol. 1975 Jan;64(1):182–210. doi: 10.1083/jcb.64.1.182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Castle J. D., Palade G. E. Secretion granules of the rabbit parotid. Selective removal of secretory contaminants from granule membranes. J Cell Biol. 1978 Feb;76(2):323–340. doi: 10.1083/jcb.76.2.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Castro Costa M. R., Edelstein S. B., Castiglione C. M., Chao H., Breakefield X. O. Properties of monoamine oxidase in control and Lesch-Nyhan fibroblasts. Biochem Genet. 1980 Jun;18(5-6):577–590. doi: 10.1007/BF00484403. [DOI] [PubMed] [Google Scholar]
  10. Durham J. P., Galanti N., Revis N. W. The purification and characterization of plasma membranes and the subcellular distribution of adenylate cyclase in mouse parotid gland. Biochim Biophys Acta. 1975 Jul 3;394(3):388–405. doi: 10.1016/0005-2736(75)90292-8. [DOI] [PubMed] [Google Scholar]
  11. Elce J. S., Broxmeyer B. Gamma-glutamyltransferase of rat kidney. Simultaneous assay of the hydrolysis and transfer reactions with (glutamate-14C)glutathione. Biochem J. 1976 Feb 1;153(2):223–232. doi: 10.1042/bj1530223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Emmelot P., Bos C. J. Studies on plasma membranes. II. K+-dependent p-nitrophenyl phosphatase activity of plasma membranes isolated from rat liver. Biochim Biophys Acta. 1966 Jun 29;121(2):375–385. [PubMed] [Google Scholar]
  13. FINDLAY J., LEVVY G. A., MARSH C. A. Inhibition of glycosidases by aldonolactones of corresponding configuration. 2. Inhibitors of beta-N-acetylglucosaminidase. Biochem J. 1958 Jul;69(3):467–476. doi: 10.1042/bj0690467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fleischer B. Isolation and characterization of Golgi apparatus and membranes from rat liver. Methods Enzymol. 1974;31:180–191. doi: 10.1016/0076-6879(74)31020-8. [DOI] [PubMed] [Google Scholar]
  15. Forte J. G., Forte G. M., Saltman P. K+-stimulated phosphatase of microsomes from gastric mucosa. J Cell Physiol. 1967 Jun;69(3):293–304. doi: 10.1002/jcp.1040690305. [DOI] [PubMed] [Google Scholar]
  16. Garrett J. R., Parsons P. A. Alkaline phosphatase and myoepithelial cells in the parotid gland of the rat. Histochem J. 1973 Sep;5(5):463–471. doi: 10.1007/BF01012003. [DOI] [PubMed] [Google Scholar]
  17. Grynszpan-Winograd O. Morphological aspects of exocytosin in the adrenal medulla. Philos Trans R Soc Lond B Biol Sci. 1971 Jun 17;261(839):291–292. doi: 10.1098/rstb.1971.0058. [DOI] [PubMed] [Google Scholar]
  18. HANES C. S., HIRD F. J. R. Synthesis of peptides in enzymic reactions involving glutathione. Nature. 1950 Aug 19;166(4216):288–292. doi: 10.1038/166288a0. [DOI] [PubMed] [Google Scholar]
  19. Hand A. R. Morphology and cytochemistry of the Golgi apparatus of rat salivary gland acnar cells. Am J Anat. 1971 Feb;130(2):141–157. doi: 10.1002/aja.1001300203. [DOI] [PubMed] [Google Scholar]
  20. Hata K., Abiko Y., Takiguchi H. Species distribution of gamma-glutamyl transpeptidase in the parotid gland. J Dent Res. 1980 Apr;59(4):728–728. doi: 10.1177/00220345800590041401. [DOI] [PubMed] [Google Scholar]
  21. Howell K. E., Ito A., Palade G. E. Endoplasmic reticulum marker enzymes in Golgi fractions--what does this mean? J Cell Biol. 1978 Nov;79(2 Pt 1):581–589. doi: 10.1083/jcb.79.2.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. ICHIKAWA A. FINE STRUCTURAL CHANGES IN RESPONSE TO HORMONAL STIMULATION OF THE PERFUSED CANINE PANCREAS. J Cell Biol. 1965 Mar;24:369–385. doi: 10.1083/jcb.24.3.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jarasch E. D., Kartenbeck J., Bruder G., Fink A., Morré D. J., Franke W. W. B-type cytochromes in plasma membranes isolated from rat liver, in comparison with those of endomembranes. J Cell Biol. 1979 Jan;80(1):37–52. doi: 10.1083/jcb.80.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kim Y. S., Perdomo J., Nordberg J. Glycoprortein biosynthesis in small intestinal mucosa. I. A study of glycosyltransferases in microsomal subfractions. J Biol Chem. 1971 Sep 10;246(17):5466–5476. [PubMed] [Google Scholar]
  25. Kinsey W. H., Decker G. L., Lennarz W. J. Isolation and partial characterization of the plasma membrane of the sea urchin egg. J Cell Biol. 1980 Oct;87(1):248–254. doi: 10.1083/jcb.87.1.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lawson D., Raff M. C., Gomperts B., Fewtrell C., Gilula N. B. Molecular events during membrane fusion. A study of exocytosis in rat peritoneal mast cells. J Cell Biol. 1977 Feb;72(2):242–259. doi: 10.1083/jcb.72.2.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Marathe G. V., Nash B., Haschemeyer R. H., Tate S. S. Ultrastructural localization of gamma-glutamyl transpeptidase in rat kidney and jejunum. FEBS Lett. 1979 Nov 15;107(2):436–440. doi: 10.1016/0014-5793(79)80425-1. [DOI] [PubMed] [Google Scholar]
  28. Max E. E., Goodman D. B., Rasmussen H. Purification and characterization of chick intestine brush border membrane. Effects of 1alpha(OH) vitamin D3 treatment. Biochim Biophys Acta. 1978 Aug 4;511(2):224–239. doi: 10.1016/0005-2736(78)90316-4. [DOI] [PubMed] [Google Scholar]
  29. Meister A., Tate S. S. Glutathione and related gamma-glutamyl compounds: biosynthesis and utilization. Annu Rev Biochem. 1976;45:559–604. doi: 10.1146/annurev.bi.45.070176.003015. [DOI] [PubMed] [Google Scholar]
  30. Meldolesi J., Jamieson J. D., Palade G. E. Composition of cellular membranes in the pancreas of the guinea pig. II. Lipids. J Cell Biol. 1971 Apr;49(1):130–149. doi: 10.1083/jcb.49.1.130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. NACHLAS M. M., MONIS B., ROSENBATT D., SELIGMAN A. M. Improvement in the histochemical localization of leucine aminopeptidase with a new substrate, L-leucyl-4-methoxy-2-naphthylamide. J Biophys Biochem Cytol. 1960 Apr;7:261–264. doi: 10.1083/jcb.7.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. PARKS H. F. On the fine structure of the parotid gland of mouse and rat. Am J Anat. 1961 May;108:303–329. doi: 10.1002/aja.1001080306. [DOI] [PubMed] [Google Scholar]
  33. Palade G. Intracellular aspects of the process of protein synthesis. Science. 1975 Aug 1;189(4200):347–358. doi: 10.1126/science.1096303. [DOI] [PubMed] [Google Scholar]
  34. Peters T. J., Müller M., De Duve C. Lysosomes of the arterial wall. I. Isolation and subcellular fractionation of cells from normal rabbit aorta. J Exp Med. 1972 Nov 1;136(5):1117–1139. doi: 10.1084/jem.136.5.1117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pratten M. K., Williams M. A., Cope G. H. Compartmentation of enzymes in the rabbit parotid salivary gland. A study by enzyme histochemical, tissue fractionation and morphometric techniques. Histochem J. 1977 Sep;9(5):573–598. doi: 10.1007/BF01002903. [DOI] [PubMed] [Google Scholar]
  36. Rutenburg A. M., Kim H., Fischbein J. W., Hanker J. S., Wasserkrug H. L., Seligman A. M. Histochemical and ultrastructural demonstration of gamma-glutamyl transpeptidase activity. J Histochem Cytochem. 1969 Aug;17(8):517–526. doi: 10.1177/17.8.517. [DOI] [PubMed] [Google Scholar]
  37. SCHRAMM M., BEN-ZVI R., BDOLAH A. EPINEPHRIN-ACTIVATED AMYLASE SECRETION IN PAROTID SLICES AND LEAKAGE OF THE ENZYME IN THE COLD. Biochem Biophys Res Commun. 1965 Feb 3;18:446–451. doi: 10.1016/0006-291x(65)90729-1. [DOI] [PubMed] [Google Scholar]
  38. SCHRAMM M., DANON D. The mechanism of enzyme secretion by the cell. I. Storage of amylase in the zymogen granules of the rat-parotis gland. Biochim Biophys Acta. 1961 Jun 10;50:102–112. doi: 10.1016/0006-3002(61)91065-4. [DOI] [PubMed] [Google Scholar]
  39. Schultz G. S., Sarras M. P., Jr, Gunther G. R., Hull B. E., Alicea H. A., Gorelick F. S., Jamieson J. D. Guinea pig pancreatic acini prepared with purified collagenase. Exp Cell Res. 1980 Nov;130(1):49–62. doi: 10.1016/0014-4827(80)90041-5. [DOI] [PubMed] [Google Scholar]
  40. Sottocasa G. L., Kuylenstierna B., Ernster L., Bergstrand A. An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J Cell Biol. 1967 Feb;32(2):415–438. doi: 10.1083/jcb.32.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Specian R. D., Neutra M. R. Mechanism of rapid mucus secretion in goblet cells stimulated by acetylcholine. J Cell Biol. 1980 Jun;85(3):626–640. doi: 10.1083/jcb.85.3.626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Stein Y., Widnell C., Stein O. Acylation of lysophosphatides by plasma membrane fractions of rat liver. J Cell Biol. 1968 Oct;39(1):185–192. doi: 10.1083/jcb.39.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tate S. S., Meister A. Interaction of gamma-glutamyl transpeptidase with amino acids, dipeptides, and derivatives and analogs of glutathione. J Biol Chem. 1974 Dec 10;249(23):7593–7602. [PubMed] [Google Scholar]
  44. Tate S. S., Meister A. Serine-borate complex as a transition-state inhibitor of gamma-glutamyl transpeptidase. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4806–4809. doi: 10.1073/pnas.75.10.4806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Touster O., Aronson N. N., Jr, Dulaney J. T., Hendrickson H. Isolation of rat liver plasma membranes. Use of nucleotide pyrophosphatase and phosphodiesterase I as marker enzymes. J Cell Biol. 1970 Dec;47(3):604–618. doi: 10.1083/jcb.47.3.604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wallach D., Kirshner N., Schramm M. Non-parallel transport of membrane proteins and content proteins during assembly of the secretory granule in rat parotid gland. Biochim Biophys Acta. 1975 Jan 14;375(1):87–105. doi: 10.1016/0005-2736(75)90074-7. [DOI] [PubMed] [Google Scholar]
  47. Widnell C. C., Unkeless J. C. Partial purification of a lipoprotein with 5'-nucleotidase activity from membranes of rat liver cells. Proc Natl Acad Sci U S A. 1968 Nov;61(3):1050–1057. doi: 10.1073/pnas.61.3.1050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Williams M. A., Pratten M. K., Turner J. W., Cope G. H. Comparative studies on the nature of purified cytomembranes of the rabbit parotid gland. Histochem J. 1979 Jan;11(1):19–50. doi: 10.1007/BF01041264. [DOI] [PubMed] [Google Scholar]
  49. Wootton A. M., Neale G., Moss D. W. Enzyme activities of cells of different types isolated from livers of normal and cholestatic rats. Clin Sci Mol Med. 1977 Jun;52(6):585–590. doi: 10.1042/cs0520585. [DOI] [PubMed] [Google Scholar]
  50. Yamashina S., Kawai K. Cytochemical studies on the localization of 5'-nucleotidase in the acinar cells of the rat salivary glands. Histochemistry. 1979 Apr 12;60(3):255–263. doi: 10.1007/BF00500655. [DOI] [PubMed] [Google Scholar]
  51. Zinder O., Hoffman P. G., Bonner W. M., Pollard H. B. Comparison of chemical properties of purified plasma membranes and secretory vesicle membranes from bovine adrenal medulla. Cell Tissue Res. 1978 Apr 17;188(2):153–170. doi: 10.1007/BF00222628. [DOI] [PubMed] [Google Scholar]

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