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
. 1988 Aug;85(15):5566–5570. doi: 10.1073/pnas.85.15.5566

Alveolar type II cells express a high-affinity receptor for pulmonary surfactant protein A.

Y Kuroki 1, R J Mason 1, D R Voelker 1
PMCID: PMC281799  PMID: 2840667

Abstract

Primary cultures of rat alveolar type II cells bind radiolabeled pulmonary surfactant protein A (SP-A) with high affinity. The binding of 125I-labeled SP-A is time- and temperature-dependent and is not accompanied by significant degradation. The binding process is saturable at low concentrations of SP-A (5 micrograms/ml), and unlabeled SP-A readily competes with labeled SP-A for cellular binding sites. Subsequent to binding, two pools of cell-associated 125I-labeled SP-A can be identified based upon sensitivity to trypsin at 0 degrees C. It is likely that the trypsin-sensitive pool comprises 125I-labeled SP-A bound to the cell surface and the trypsin-insensitive pool comprises the internalized protein. Scatchard analysis of cell surface binding of SP-A at 0.1-10 micrograms/ml shows positive cooperativity at concentrations between 0.1 and 1 micrograms/ml. Hill plots give nH = 1.34 +/- 0.08 with an apparent dissociation constant K'd = 1.02 +/- 0.32 micrograms/ml (which is 0.64 +/- 0.19 nM if the native molecular mass of oligomeric SP-A is assumed to be 1.6 MDa). The binding of SP-A to type II cells shows an absolute requirement for Ca2+. The putative receptor for SP-A is unaffected by treatment of type II cells with a variety of proteases and N-Glycanase (EC 3.5.1.52). Alveolar macrophages also exhibit high-affinity binding of SP-A, but rat lung fibroblasts and the alveolar epithelial cell line L2 exhibit only nonspecific binding.

Full text

PDF
5566

Images in this article

5567Image
on p.5567
5567Image
on p.5567

Selected References

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

  1. Benson B., Hawgood S., Schilling J., Clements J., Damm D., Cordell B., White R. T. Structure of canine pulmonary surfactant apoprotein: cDNA and complete amino acid sequence. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6379–6383. doi: 10.1073/pnas.82.19.6379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Claypool W. D., Wang D. L., Chander A., Fisher A. B. An ethanol/ether soluble apoprotein from rat lung surfactant augments liposome uptake by isolated granular pneumocytes. J Clin Invest. 1984 Sep;74(3):677–684. doi: 10.1172/JCI111483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dautry-Varsat A., Ciechanover A., Lodish H. F. pH and the recycling of transferrin during receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2258–2262. doi: 10.1073/pnas.80.8.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dethloff L. A., Gilmore L. B., Brody A. R., Hook G. E. Induction of intra- and extra-cellular phospholipids in the lungs of rats exposed to silica. Biochem J. 1986 Jan 1;233(1):111–118. doi: 10.1042/bj2330111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dobbs L. G., Mason R. J. Pulmonary alveolar type II cells isolated from rats. Release of phosphatidylcholine in response to beta-adrenergic stimulation. J Clin Invest. 1979 Mar;63(3):378–387. doi: 10.1172/JCI109313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dobbs L. G., Wright J. R., Hawgood S., Gonzalez R., Venstrom K., Nellenbogen J. Pulmonary surfactant and its components inhibit secretion of phosphatidylcholine from cultured rat alveolar type II cells. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1010–1014. doi: 10.1073/pnas.84.4.1010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Douglas W. H., Kaighn M. E. Clonal isolation of differentiated rat lung cells. In Vitro. 1974 Sep-Oct;10(3-4):230–237. doi: 10.1007/BF02615237. [DOI] [PubMed] [Google Scholar]
  9. Fanestil D. D., Barrows C. H., Jr Aging in the rotifer. J Gerontol. 1965 Oct;20(4):462–469. [PubMed] [Google Scholar]
  10. Floros J., Phelps D. S., Taeusch H. W. Biosynthesis and in vitro translation of the major surfactant-associated protein from human lung. J Biol Chem. 1985 Jan 10;260(1):495–500. [PubMed] [Google Scholar]
  11. Floros J., Steinbrink R., Jacobs K., Phelps D., Kriz R., Recny M., Sultzman L., Jones S., Taeusch H. W., Frank H. A. Isolation and characterization of cDNA clones for the 35-kDa pulmonary surfactant-associated protein. J Biol Chem. 1986 Jul 5;261(19):9029–9033. [PubMed] [Google Scholar]
  12. Fornstedt N., Porath J. Characterization studies on a new lectin found in seeds of Vicia ervilia. FEBS Lett. 1975 Sep 15;57(2):187–191. doi: 10.1016/0014-5793(75)80713-7. [DOI] [PubMed] [Google Scholar]
  13. Glasser S. W., Korfhagen T. R., Weaver T., Pilot-Matias T., Fox J. L., Whitsett J. A. cDNA and deduced amino acid sequence of human pulmonary surfactant-associated proteolipid SPL(Phe). Proc Natl Acad Sci U S A. 1987 Jun;84(12):4007–4011. doi: 10.1073/pnas.84.12.4007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goldstein J. L., Basu S. K., Brown M. S. Receptor-mediated endocytosis of low-density lipoprotein in cultured cells. Methods Enzymol. 1983;98:241–260. doi: 10.1016/0076-6879(83)98152-1. [DOI] [PubMed] [Google Scholar]
  15. Goldstein J. L., Brown M. S. Binding and degradation of low density lipoproteins by cultured human fibroblasts. Comparison of cells from a normal subject and from a patient with homozygous familial hypercholesterolemia. J Biol Chem. 1974 Aug 25;249(16):5153–5162. [PubMed] [Google Scholar]
  16. Haagsman H. P., Hawgood S., Sargeant T., Buckley D., White R. T., Drickamer K., Benson B. J. The major lung surfactant protein, SP 28-36, is a calcium-dependent, carbohydrate-binding protein. J Biol Chem. 1987 Oct 15;262(29):13877–13880. [PubMed] [Google Scholar]
  17. Hawgood S., Benson B. J., Hamilton R. L., Jr Effects of a surfactant-associated protein and calcium ions on the structure and surface activity of lung surfactant lipids. Biochemistry. 1985 Jan 1;24(1):184–190. doi: 10.1021/bi00322a026. [DOI] [PubMed] [Google Scholar]
  18. Hawgood S., Benson B. J., Schilling J., Damm D., Clements J. A., White R. T. Nucleotide and amino acid sequences of pulmonary surfactant protein SP 18 and evidence for cooperation between SP 18 and SP 28-36 in surfactant lipid adsorption. Proc Natl Acad Sci U S A. 1987 Jan;84(1):66–70. doi: 10.1073/pnas.84.1.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Innerarity T. L., Pitas R. E., Mahley R. W. Lipoprotein-receptor interactions. Methods Enzymol. 1986;129:542–565. doi: 10.1016/0076-6879(86)29091-6. [DOI] [PubMed] [Google Scholar]
  20. Jacobs H. C., Ikegami M., Jobe A. H., Berry D. D., Jones S. Reutilization of surfactant phosphatidylcholine in adult rabbits. Biochim Biophys Acta. 1985 Oct 23;837(1):77–84. doi: 10.1016/0005-2760(85)90087-6. [DOI] [PubMed] [Google Scholar]
  21. Jacobs H., Jobe A., Ikegami M., Conaway D. The significance of reutilization of surfactant phosphatidylcholine. J Biol Chem. 1983 Apr 10;258(7):4159–4165. [PubMed] [Google Scholar]
  22. Jacobs H., Jobe A., Ikegami M., Jones S. Surfactant phosphatidylcholine source, fluxes, and turnover times in 3-day-old, 10-day-old, and adult rabbits. J Biol Chem. 1982 Feb 25;257(4):1805–1810. [PubMed] [Google Scholar]
  23. King R. J., Klass D. J., Gikas E. G., Clements J. A. Isolation of apoproteins from canine surface active material. Am J Physiol. 1973 Apr;224(4):788–795. doi: 10.1152/ajplegacy.1973.224.4.788. [DOI] [PubMed] [Google Scholar]
  24. Kuroki Y., Mason R. J., Voelker D. R. Pulmonary surfactant apoprotein A structure and modulation of surfactant secretion by rat alveolar type II cells. J Biol Chem. 1988 Mar 5;263(7):3388–3394. [PubMed] [Google Scholar]
  25. Rice W. R., Ross G. F., Singleton F. M., Dingle S., Whitsett J. A. Surfactant-associated protein inhibits phospholipid secretion from type II cells. J Appl Physiol (1985) 1987 Aug;63(2):692–698. doi: 10.1152/jappl.1987.63.2.692. [DOI] [PubMed] [Google Scholar]
  26. Ross G. F., Notter R. H., Meuth J., Whitsett J. A. Phospholipid binding and biophysical activity of pulmonary surfactant-associated protein (SAP)-35 and its non-collagenous COOH-terminal domains. J Biol Chem. 1986 Oct 25;261(30):14283–14291. [PubMed] [Google Scholar]
  27. Sano K., Fisher J., Mason R. J., Kuroki Y., Schilling J., Benson B., Voelker D. Isolation and sequence of a cDNA clone for the rat pulmonary surfactant-associated protein (PSP-A). Biochem Biophys Res Commun. 1987 Apr 14;144(1):367–374. doi: 10.1016/s0006-291x(87)80519-3. [DOI] [PubMed] [Google Scholar]
  28. Van Heyningen W. E., Carpenter C. C., Pierce N. F., Greenough W. B., 3rd Deactivation of cholera toxin by ganglioside. J Infect Dis. 1971 Oct;124(4):415–418. doi: 10.1093/infdis/124.4.415. [DOI] [PubMed] [Google Scholar]
  29. Walker S. R., Williams M. C., Benson B. Immunocytochemical localization of the major surfactant apoproteins in type II cells, Clara cells, and alveolar macrophages of rat lung. J Histochem Cytochem. 1986 Sep;34(9):1137–1148. doi: 10.1177/34.9.2426341. [DOI] [PubMed] [Google Scholar]
  30. Weaver T. E., Hull W. M., Ross G. F., Whitsett J. A. Intracellular and oligomeric forms of surfactant-associated apolipoproteins(s) A in the rat. Biochim Biophys Acta. 1985 Mar 1;827(3):260–267. doi: 10.1016/0167-4838(85)90210-9. [DOI] [PubMed] [Google Scholar]
  31. Whitsett J. A., Weaver T., Hull W., Ross G., Dion C. Synthesis of surfactant-associated glycoprotein A by rat type II epithelial cells. Primary translation products and post-translational modification. Biochim Biophys Acta. 1985 Apr 5;828(2):162–171. doi: 10.1016/0167-4838(85)90053-6. [DOI] [PubMed] [Google Scholar]
  32. Williams M. C., Benson B. J. Immunocytochemical localization and identification of the major surfactant protein in adult rat lung. J Histochem Cytochem. 1981 Feb;29(2):291–305. doi: 10.1177/29.2.7019304. [DOI] [PubMed] [Google Scholar]
  33. Willingham M. C., Pastan I. Ultrastructural immunocytochemical localization of the transferrin receptor using a monoclonal antibody in human KB cells. J Histochem Cytochem. 1985 Jan;33(1):59–64. doi: 10.1177/33.1.2856926. [DOI] [PubMed] [Google Scholar]
  34. Wright J. R., Wager R. E., Hawgood S., Dobbs L., Clements J. A. Surfactant apoprotein Mr = 26,000-36,000 enhances uptake of liposomes by type II cells. J Biol Chem. 1987 Feb 25;262(6):2888–2894. [PubMed] [Google Scholar]
  35. Yamashiro D. J., Tycko B., Fluss S. R., Maxfield F. R. Segregation of transferrin to a mildly acidic (pH 6.5) para-Golgi compartment in the recycling pathway. Cell. 1984 Jul;37(3):789–800. doi: 10.1016/0092-8674(84)90414-8. [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