• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. Nov 15, 1995; 312(Pt 1): 175–181.
PMCID: PMC1136242

Ca(2+)-dependent binding of annexin IV to surfactant protein A and lamellar bodies in alveolar type II cells.


Surfactant protein A (SP-A), a lung-specific glycoprotein in pulmonary surfactant, is synthesized and secreted from the alveolar type II cells. It has been shown that SP-A is a Ca(2+)-binding protein with several binding sites and that the high-affinity site(s) is located in the C-terminal region of SP-A. In the present study we isolated the proteins from bovine lung soluble fraction that bind to SP-A in a Ca(2+)-dependent manner using DEAE-Sephacel and SP-A-conjugated Sepharose 4B. At least three different protein bands with molecular masses of 24.5, 32, and 33 kDa were observed on SDS/PAGE. The main protein, with molecular mass of 32 kDa, was identified as annexin IV by the partial-amino-acid-sequence analyses and an immunoblot analysis with anti-(annexin IV) antiserum. We also found from the immunoblot analysis that the cytosolic fraction of isolated rat alveolar type II cells contains annexin IV. In addition, when rat lung cytosol was loaded on to the lung lamellar body-conjugated Sepharose 4B in the presence of Ca2+, two proteins, with molecular masses of 32 and 60 kDa on SDS/PAGE respectively, were eluted with EGTA. The 32 kDa protein was shown to be annexin IV by an immunoblot analysis with the antiserum against annexin IV. The lung annexin IV augmented the Ca(2+)-induced aggregation of the lung lamellar bodies from rats. However, the augmentation of aggregation of the lung lamellar bodies by annexin IV was attenuated when the lamellar bodies were preincubated with polyclonal anti-SP-A antibodies. SP-A bound to annexin IV under conditions where contaminated lipid was removed. These results suggest that SP-A bound to annexin IV based on protein-protein interaction, though both proteins are phospholipid-binding proteins. All these findings suggest that the interaction between SP-A and annexin IV may have some role in alveolar type II cells.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • King RJ, Clements JA. Surface active materials from dog lung. I. Method of isolation. Am J Physiol. 1972 Sep;223(3):707–714. [PubMed]
  • King RJ, Klass DJ, Gikas EG, Clements JA. Isolation of apoproteins from canine surface active material. Am J Physiol. 1973 Apr;224(4):788–795. [PubMed]
  • White RT, Damm D, Miller J, Spratt K, Schilling J, Hawgood S, Benson B, Cordell B. Isolation and characterization of the human pulmonary surfactant apoprotein gene. Nature. 317(6035):361–363. [PubMed]
  • Ross GF, Notter RH, Meuth J, Whitsett JA. 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]
  • Sano K, Fisher J, Mason RJ, 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. [PubMed]
  • Boggaram V, Qing K, Mendelson CR. The major apoprotein of rabbit pulmonary surfactant. Elucidation of primary sequence and cyclic AMP and developmental regulation. J Biol Chem. 1988 Feb 25;263(6):2939–2947. [PubMed]
  • Kuroki Y, Akino T. Pulmonary surfactant protein A (SP-A) specifically binds dipalmitoylphosphatidylcholine. J Biol Chem. 1991 Feb 15;266(5):3068–3073. [PubMed]
  • Haagsman HP, White RT, Schilling J, Lau K, Benson BJ, Golden J, Hawgood S, Clements JA. Studies of the structure of lung surfactant protein SP-A. Am J Physiol. 1989 Dec;257(6 Pt 1):L421–L429. [PubMed]
  • De Lisle RC, Williams JA. Regulation of membrane fusion in secretory exocytosis. Annu Rev Physiol. 1986;48:225–238. [PubMed]
  • Haagsman HP, Sargeant T, Hauschka PV, Benson BJ, Hawgood S. Binding of calcium to SP-A, a surfactant-associated protein. Biochemistry. 1990 Sep 25;29(38):8894–8900. [PubMed]
  • Sohma H, Watanabe T, Kuroki Y, Yoshino H, Matsushima N, Yazawa M, Akino T. Calcium dependent conformational changes of surfactant protein A (SP-A) and its collagenase resistant fragment with or without dithiothreitol. Biochim Biophys Acta. 1992 Sep 23;1159(2):150–154. [PubMed]
  • Sohma H, Hattori A, Kuroki Y, Akino T. Calcium and dithiothreitol dependent conformational changes in beta-sheet structure of collagenase resistant fragment of human surfactant protein A. Biochem Mol Biol Int. 1993 Jun;30(2):329–336. [PubMed]
  • Suzuki Y, Fujita Y, Kogishi K. Reconstitution of tubular myelin from synthetic lipids and proteins associated with pig pulmonary surfactant. Am Rev Respir Dis. 1989 Jul;140(1):75–81. [PubMed]
  • Hawgood S, Benson BJ, Hamilton RL., 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. [PubMed]
  • Kuroki Y, Mason RJ, Voelker DR. Alveolar type II cells express a high-affinity receptor for pulmonary surfactant protein A. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5566–5570. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Duck-Chong CG. The isolation of lamellar bodies and their membranous content from rat lung, lamb tracheal fluid and human amniotic fluid. Life Sci. 1978 Jun 12;22(22):2025–2030. [PubMed]
  • Kohn J, Wilchek M. A new approach (cyano-transfer) for cyanogen bromide activation of Sepharose at neutral pH, which yields activated resins, free of interfering nitrogen derivatives. Biochem Biophys Res Commun. 1982 Aug;107(3):878–884. [PubMed]
  • Creutz CE. Secretory vesicle - cytosol interactions in exocytosis: isolation by Ca2+-dependent affinity chromatography of proteins that bind to the chromaffin granule membrane. Biochem Biophys Res Commun. 1981 Dec 31;103(4):1395–1400. [PubMed]
  • Dobbs LG, Mason RJ. 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. [PMC free article] [PubMed]
  • Sohma H, Hashimoto H, Ohguro H, Akino T. Two gamma-subunits, gamma-I and gamma-II, complex with the same beta-subunits in bovine brain G-proteins (Gi/o). Biochem Biophys Res Commun. 1992 Apr 15;184(1):175–182. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • BARTLETT GR. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed]
  • Phizackerley PJ, Town MH, Newman GE. Hydrophobic proteins of lamellated osmiophilic bodies isolated from pig lung. Biochem J. 1979 Dec 1;183(3):731–736. [PMC free article] [PubMed]
  • Creutz CE. The annexins and exocytosis. Science. 1992 Nov 6;258(5084):924–931. [PubMed]
  • Creutz CE, Zaks WJ, Hamman HC, Crane S, Martin WH, Gould KL, Oddie KM, Parsons SJ. Identification of chromaffin granule-binding proteins. Relationship of the chromobindins to calelectrin, synhibin, and the tyrosine kinase substrates p35 and p36. J Biol Chem. 1987 Feb 5;262(4):1860–1868. [PubMed]
  • Kuroki Y, Fukada Y, Takahashi H, Akino T. Monoclonal antibodies against human pulmonary surfactant apoproteins: specificity and application in immunoassay. Biochim Biophys Acta. 1985 Sep 11;836(2):201–209. [PubMed]
  • Oosterlaken-Dijksterhuis MA, van Eijk M, van Buel BL, van Golde LM, Haagsman HP. Surfactant protein composition of lamellar bodies isolated from rat lung. Biochem J. 1991 Feb 15;274(Pt 1):115–119. [PMC free article] [PubMed]
  • Hawgood S, Benson BJ, Hamilton RL., 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. [PubMed]
  • Dobbs LG, Wright JR, 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. [PMC free article] [PubMed]
  • Rice WR, Ross GF, Singleton FM, Dingle S, Whitsett JA. Surfactant-associated protein inhibits phospholipid secretion from type II cells. J Appl Physiol (1985) 1987 Aug;63(2):692–698. [PubMed]
  • Wright JR, Borchelt JD, Hawgood S. Lung surfactant apoprotein SP-A (26-36 kDa) binds with high affinity to isolated alveolar type II cells. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5410–5414. [PMC free article] [PubMed]
  • Coalson JJ, Winter VT, Martin HM, King RJ. Colloidal gold immunoultrastructural localization of rat surfactant. Am Rev Respir Dis. 1986 Feb;133(2):230–237. [PubMed]
  • Junker M, Creutz CE. Ca(2+)-dependent binding of endonexin (annexin IV) to membranes: analysis of the effects of membrane lipid composition and development of a predictive model for the binding interaction. Biochemistry. 1994 Aug 2;33(30):8930–8940. [PubMed]
  • Montfoort A, van Golde LM, van Deenen LL. Molecular species of lecithins from various animal tissues. Biochim Biophys Acta. 1971 Mar 16;231(2):335–342. [PubMed]
  • Adachi H, Hayashi H, Sato H, Dempo K, Akino T. Characterization of phospholipids accumulated in pulmonary-surfactant compartments of rats intratracheally exposed to silica. Biochem J. 1989 Sep 15;262(3):781–786. [PMC free article] [PubMed]
  • Creutz CE. cis-Unsaturated fatty acids induce the fusion of chromaffin granules aggregated by synexin. J Cell Biol. 1981 Oct;91(1):247–256. [PMC free article] [PubMed]
  • Hamman HC, Gaffey LC, Lynch KR, Creutz CE. Cloning and characterization of a cDNA encoding bovine endonexin (chromobindin 4). Biochem Biophys Res Commun. 1988 Oct 31;156(2):660–667. [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...