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. 2010 Feb 6;1(2):188–197. doi: 10.1007/s13238-010-0024-z

Surfactant protein A (SP-A) binds to phosphatidylserine and competes with annexin V binding on late apoptotic cells

Anne Jäkel 1,✉, Kenneth B M Reid 2, Howard Clark 3
PMCID: PMC4875166  PMID: 21203987

Abstract

The role of surfactant protein A (SP-A) in the recognition and clearance of apoptotic cells is well established, but to date, it is still not clear which surface molecules of apoptotic cells are involved in the process. Here we present evidence that phosphatidylserine (PS) is a relevant binding molecule for human SP-A. The binding is Ca2+-dependent and is not inhibited by mannose, suggesting that the sugar-binding site of the carbohydrate recognition domain (CRD) of SP-A is not involved. Flow cytometry studies on apoptotic Jurkat cells revealed apparent inhibition of annexin V binding by increasing concentrations of SP-A in late apoptotic but not early apoptotic cells, and this was consistent for Jurkat cells and neutrophils. Supporting these data, confocal microscopy results show a co-localisation of annexin V and SP-A in late apoptotic but not early apoptotic cells. However, we cannot conclude that this inhibition is exclusively due to the binding of SP-A to PS on the cell surface, as annexin V is not wholly specific for PS and SP-A also interacts with other phospholipids that might become exposed on the apoptotic cell surface.

Keywords: surfactant proteins, phosphatidylserine, apoptotic cells

References

  1. Bennett M.R., Gibson D.F., Schwartz S.M., Tait J.F. Binding and phagocytosis of apoptotic vascular smooth muscle cells is mediated in part by exposure of phosphatidylserine. Circ Res. 1995;77:1136–1142. doi: 10.1161/01.RES.77.6.1136. [DOI] [PubMed] [Google Scholar]
  2. Casals C., Miguel E., Perez-Gil J. Tryptophan fluorescence study on the interaction of pulmonary surfactant protein A with phospholipid vesicles. Biochem J. 1993;296(Pt3):585–593. doi: 10.1042/bj2960585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clark H., Palaniyar N., Hawgood S., Reid K.B. A recombinant fragment of human surfactant protein D reduces alveolar macrophage apoptosis and pro-inflammatory cytokines in mice developing pulmonary emphysema. Ann N Y Acad Sci. 2003;1010:113–116. doi: 10.1196/annals.1299.019. [DOI] [PubMed] [Google Scholar]
  4. Clark H., Palaniyar N., Strong P., Edmondson J., Hawgood S., Reid K.B. Surfactant protein D reduces alveolar macrophage apoptosis in vivo. J Immunol. 2002;169:2892–2899. doi: 10.4049/jimmunol.169.6.2892. [DOI] [PubMed] [Google Scholar]
  5. Clark H., Reid K.B. Structural requirements for SP-D function in vitro and in vivo: therapeutic potential of recombinant SP-D. Immunobiology. 2002;205:619–631. doi: 10.1078/0171-2985-00159. [DOI] [PubMed] [Google Scholar]
  6. Crouch E.C. Collectins and pulmonary host defense. Am J Respir Cell Mol Biol. 1998;19:177–201. doi: 10.1165/ajrcmb.19.2.140. [DOI] [PubMed] [Google Scholar]
  7. Fadok V.A., de Cathelineau A., Daleke D.L., Henson P.M., Bratton D.L. Loss of phospholipid asymmetry and surface exposure of phosphatidylserine is required for phagocytosis of apoptotic cells by macrophages and fibroblasts. J Biol Chem. 2001;276:1071–1077. doi: 10.1074/jbc.M003649200. [DOI] [PubMed] [Google Scholar]
  8. Fadok V.A., Savill J.S., Haslett C., Bratton D.L., Doherty D.E., Campbell P.A., Henson P.M. Different populations of macrophages use either the vitronectin receptor or the phosphatidylserine receptor to recognize and remove apoptotic cells. J Immunol. 1992;149:4029–4035. [PubMed] [Google Scholar]
  9. Fadok V.A., Voelker D.R., Campbell P.A., Cohen J.J., Bratton D.L., Henson P.M. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J Immunol. 1992;148:2207–2216. [PubMed] [Google Scholar]
  10. Hart S.P., Ross J.A., Ross K., Haslett C., Dransfield I. Molecular characterization of the surface of apoptotic neutrophils: implications for functional downregulation and recognition by phagocytes. Cell Death Differ. 2000;7:493–503. doi: 10.1038/sj.cdd.4400680. [DOI] [PubMed] [Google Scholar]
  11. Henson P.M., Bratton D.L., Fadok V.A. The phosphatidylserine receptor: a crucial molecular switch? Nat Rev Mol Cell Biol. 2001;2:627–633. doi: 10.1038/35085094. [DOI] [PubMed] [Google Scholar]
  12. Holmskov U., Malhotra R., Sim R.B., Jensenius J.C. Collectins: collagenous C-type lectins of the innate immune defense system. Immunol Today. 1994;15:67–74. doi: 10.1016/0167-5699(94)90136-8. [DOI] [PubMed] [Google Scholar]
  13. Homburg C.H., de Haas M., von dem Borne A.E., Verhoeven A.J., Reutelingsperger C.P., Roos D. Human neutrophils lose their surface Fc gamma RIII and acquire Annexin V binding sites during apoptosis in vitro. Blood. 1995;85:532–540. [PubMed] [Google Scholar]
  14. Hoppe H.J., Reid K.B. Collectins-soluble proteins containing collagenous regions and lectin domains-and their roles in innate immunity. Protein Sci. 1994;3:1143–1158. doi: 10.1002/pro.5560030801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jakel, A., Clark, H., Reid, K.B., and Sim, R.B. (2009). The human lung surfactant proteins A (SP-A) and D (SP-D) interact with apoptotic target cells by different binding mechanisms. Immunobiology. In press. [DOI] [PubMed]
  16. Kerr J.F., Wyllie A.H., Currie A.R. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972;26:239–257. doi: 10.1038/bjc.1972.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kilpatrick D.C. Phospholipid-binding activity of human mannan-binding lectin. Immunol Lett. 1998;61:191–195. doi: 10.1016/S0165-2478(98)00031-5. [DOI] [PubMed] [Google Scholar]
  18. Kuroki Y., Akino T. Pulmonary surfactant protein A (SPA) specifically binds dipalmitoylphosphatidylcholine. J Biol Chem. 1991;266:3068–3073. [PubMed] [Google Scholar]
  19. Lu J., Teh C., Kishore U., Reid K.B. Collectins and ficolins: sugar pattern recognition molecules of the mammalian innate immune system. Biochim Biophys Acta. 2002;1572:387–400. doi: 10.1016/S0304-4165(02)00320-3. [DOI] [PubMed] [Google Scholar]
  20. Malhotra R., Lu J., Holmskov U., Sim R.B. Collectins, collectin receptors and the lectin pathway of complement activation. Clin Exp Immunol. 1994;97(Suppl2):4–9. doi: 10.1111/j.1365-2249.1994.tb06254.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Martin S.J., Reutelingsperger C.P., McGahon A.J., Rader J.A., van Schie R.C., LaFace D.M., Green D.R. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med. 1995;182:1545–1556. doi: 10.1084/jem.182.5.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Meers P., Mealy T. Calcium-dependent annexin V binding to phospholipids: stoichiometry, specificity, and the role of negative charge. Biochemistry. 1993;32:11711–11721. doi: 10.1021/bi00094a030. [DOI] [PubMed] [Google Scholar]
  23. Meers P., Mealy T. Phospholipid determinants for annexin V binding sites and the role of tryptophan 187. Biochemistry. 1994;33:5829–5837. doi: 10.1021/bi00185a022. [DOI] [PubMed] [Google Scholar]
  24. Meyboom A., Maretzki D., Stevens P.A., Hofmann K.P. Interaction of pulmonary surfactant protein A with phospholipid liposomes: a kinetic study on head group and fatty acid specificity. Biochim Biophys Acta. 1999;1441:23–35. doi: 10.1016/S1388-1981(99)00142-0. [DOI] [PubMed] [Google Scholar]
  25. Nauta A.J., Castellano G., Xu W., Woltman A.M., Borrias M.C., Daha M.R., van Kooten C., Roos A. Opsonization with C1q and mannose-binding lectin targets apoptotic cells to dendritic cells. J Immunol. 2004;173:3044–3050. doi: 10.4049/jimmunol.173.5.3044. [DOI] [PubMed] [Google Scholar]
  26. Ogden C.A., deCathelineau A., Hoffmann P.R., Bratton D., Ghebrehiwet B., Fadok V.A., Henson P.M. C1q and mannose binding lectin engagement of cell surface calreticulin and CD91 initiates macropinocytosis and uptake of apoptotic cells. J Exp Med. 2001;194:781–795. doi: 10.1084/jem.194.6.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Paidassi H., Tacnet-Delorme P., Garlatti V., Darnault C., Ghebrehiwet B., Gaboriaud C., Arlaud G.J., Frachet P. C1q binds phosphatidylserine and likely acts as a multiligand-bridging molecule in apoptotic cell recognition. J Immunol. 2008;180:2329–2338. doi: 10.4049/jimmunol.180.4.2329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schagat T.L., Wofford J.A., Wright J.R. Surfactant protein A enhances alveolar macrophage phagocytosis of apoptotic neutrophils. J Immunol. 2001;166:2727–2733. doi: 10.4049/jimmunol.166.4.2727. [DOI] [PubMed] [Google Scholar]
  29. Schlegel R.A., Stevens M., Lumley-Sapanski K., Williamson P. Altered lipid packing identifies apoptotic thymocytes. Immunol Lett. 1993;36:283–288. doi: 10.1016/0165-2478(93)90101-7. [DOI] [PubMed] [Google Scholar]
  30. Shiratsuchi A., Umeda M., Ohba Y., Nakanishi Y. Recognition of phosphatidylserine on the surface of apoptotic spermatogenic cells and subsequent phagocytosis by Sertoli cells of the rat. J Biol Chem. 1997;272:2354–2358. doi: 10.1074/jbc.272.4.2354. [DOI] [PubMed] [Google Scholar]
  31. Sim R.B., Moffatt B.E., Shaw J.M., Ferluga J. Complement control proteins and receptors: from FH to CR4. Molecular aspects of innate and adaptive immunity. Cambridge, UK: RSC Publishing; 2008. pp. 84–104. [Google Scholar]
  32. Suwabe A., Mason R.J., Voelker D.R. Calcium dependent association of surfactant protein A with pulmonary surfactant: application to simple surfactant protein A purification. Arch Biochem Biophys. 1996;327:285–291. doi: 10.1006/abbi.1996.0123. [DOI] [PubMed] [Google Scholar]
  33. Thiel S., Reid K.B. Structures and functions associated with the group of mammalian lectins containing collagen-like sequences. FEBS Lett. 1989;250:78–84. doi: 10.1016/0014-5793(89)80689-1. [DOI] [PubMed] [Google Scholar]
  34. Vandivier R.W., Ogden C.A., Fadok V.A., Hoffmann P.R., Brown K. K., Botto M., Walport M.J., Fisher J.H., Henson P.M., Greene K.E. Role of surfactant proteins A, D, and C1q in the clearance of apoptotic cells in vivo and in vitro: calreticulin and CD91 as a common collectin receptor complex. J Immunol. 2002;169:3978–3986. doi: 10.4049/jimmunol.169.7.3978. [DOI] [PubMed] [Google Scholar]
  35. Vermes I., Haanen C., Steffens-Nakken H., Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods. 1995;184:39–51. doi: 10.1016/0022-1759(95)00072-I. [DOI] [PubMed] [Google Scholar]
  36. Wallis R. Structural and functional aspects of complement activation by mannose-binding protein. Immunobiology. 2002;205:433–445. doi: 10.1078/0171-2985-00144. [DOI] [PubMed] [Google Scholar]
  37. Weis W.I., Taylor M.E., Drickamer K. The C-type lectin superfamily in the immune system. Immunol Rev. 1998;163:19–34. doi: 10.1111/j.1600-065X.1998.tb01185.x. [DOI] [PubMed] [Google Scholar]
  38. Wyllie A.H., Kerr J.F., Currie A.R. Cell death: the significance of apoptosis. Int Rev Cytol. 1980;68:251–306. doi: 10.1016/S0074-7696(08)62312-8. [DOI] [PubMed] [Google Scholar]

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