Molecular and Cellular Biochemistry

, Volume 304, Issue 1–2, pp 119–125 | Cite as

Characterization of the biochemical and biophysical properties of the phosphatidylserine receptor (PS-R) gene product

  • Nitu Tibrewal
  • Tong Liu
  • Hong Li
  • Raymond B. Birge


The PS-R gene product was originally described as a cell surface receptor that interacts with externalized phosphatidylserine (PS) on apoptotic cells, but more recent studies have shown that it plays a critical role in organ development and terminal differentiation of many cell types during embryogenesis. Despite these important developmental functions, the biochemical and molecular properties of PS-R are poorly understood. Here we have used several approaches to show that PS-R undergoes processive post-translational protein cross-linking to form covalent multimers within the nuclear compartment. Although PS-R has a potential Glu-Glu (QQ) duet that is often targeted by transglutaminase TG-2, the oligomerization of PS-R was not effected by QQ→AA mutation, or when PS-R gene product was expressed in TG-2 (-/-) fibroblasts. Pulse-chase experiments with 35 S-methionine indicates that the PS-R undergoes an initial proteolytic cleavage, followed by progressive multimerization of the monomeric subunits over time. In summary, we report here that PS-R is modified by an unusual post-translational modification, and we speculate that homomultimer of PS-R might be playing an important function as a scaffolding protein in the nucleus.


Phosphatidylserine Phagocytosis Apoptotic cells Protein Cross Linking Post Translational Modification 



We would like Dr. Jens Bose (German Research Center for Biotechnology) for discussing unpublished data. We would also like to thank Dr. Valerie Fadok and Dr. Peter Henson (National Jewish Hospital, Denver, CO) for PS-R DNA, Dr. Gerry Melino, (University of Roma) for the TG-2 (-/-) cells, Dr. Richard Flavell (Yale University) for PS-R (-/-) cells and Veera D’mello. Dr. Charles Reichman and Dr. Carolyn Suzuki for critical comments on the manuscipt. This work was supported by a grant from the Arthritis Foundation and from the UMDNJ Research Foundation to R.B. Birge and NIH NS 046593 to H. Li.


  1. 1.
    Fadok VA, Bratton DL, Rose DM, Pearson A, Ezekewitz RA, Henson PM (2000). A receptor for phosphatidylserine-specific clearance of apoptotic cells. Nature 405(6782):85–90PubMedCrossRefGoogle Scholar
  2. 2.
    Savill J, Fadok V (2000). Corpse clearance defines the meaning of cell death. Nature 407(6805):784–788PubMedCrossRefGoogle Scholar
  3. 3.
    Ravichandran KS (2003) “Recruitment signals” from apoptotic cells: invitation to a quiet meal. Cell 113(7):817–820PubMedCrossRefGoogle Scholar
  4. 4.
    Wu Y, Tibrewal N, Birge RB (2006) Phosphatidylserine recognition by phagocytes:a view to a kill. Trends Cell Biol 16(4):189–197PubMedCrossRefGoogle Scholar
  5. 5.
    Hanayama R, Tanaka M, Miwa K, Shinohara A, Iwamatsu A, Nagata S (2002) Identification of a factor that links apoptotic cells to phagocytes. Nature 417(6885):182–187PubMedCrossRefGoogle Scholar
  6. 6.
    Hanayama R, Tanaka M, Miwa K, Nagata S (2004) Expression of developmental endothelial locus-1 in a subset of macrophages for engulfment of apoptotic cells. J Immunol 172(6):3876–3882PubMedGoogle Scholar
  7. 7.
    Ishimoto Y, Ohashi K, Mizuno K, Nakano T (2000) Promotion of the uptake of PS liposomes and apoptotic cells by a product of growth arrest-specific gene, gas6. J Biochem (Tokyo) 127(3):411–417Google Scholar
  8. 8.
    Hall MO, Obin MS, Prieto AL, Burgess BL, Abrams TA (2002) Gas6 binding to photoreceptor outer segments requires gamma-carboxyglutamic acid (Gla) and Ca(2+) and is required for OS phagocytosis by RPE cells in vitro. Exp Eye Res 75(4):391–400PubMedCrossRefGoogle Scholar
  9. 9.
    Cikala M, Alexandrova O, David CN, Proschel M, Stiening B, Cramer P, Bottger A (2004) The phosphatidylserine receptor from Hydra is a nuclear protein with potential Fe(II) dependent oxygenase activity. BMC Cell Biol 5:26PubMedCrossRefGoogle Scholar
  10. 10.
    Cui P, Qin B, Liu N, Pan G, Pei D (2004) Nuclear localization of the phosphatidylserine receptor protein via multiple nuclear localization signals. Exp Cell Res 293(1):154–163PubMedCrossRefGoogle Scholar
  11. 11.
    Mitchell JE, Cvetanovic M, Tibrewal N, Patel V, Colamonici OR, Li MO, Flavell RA, Levine JS, Birge RB, Ucker DS (2006) The presumptive phosphatidylserine receptor is dispensable for innate anti-inflammatory recognition and clearance of apoptotic cells. J Biol Chem 281(9):5718–5725PubMedCrossRefGoogle Scholar
  12. 12.
    Wang X, Wu YC, Fadok VA, Lee MC, K. Gengyo-Ando, Cheng LC, Ledwich D, Hsu PK, Chen JY, Chou BK, Henson P, Mitani S, Xue D (2003) Cell corpse engulfment mediated by C. elegans phosphatidylserine receptor through CED-5 and CED-12. Science 302(5650):1563–1566PubMedCrossRefGoogle Scholar
  13. 13.
    Yin J, Haney L, Walk S, Zhou S, Ravichandran KS, Wang W (2004) Nuclear localization of the DOCK180/ELMO complex. Arch Biochem Biophys 429(1):23–29PubMedCrossRefGoogle Scholar
  14. 14.
    Bose J, Gruber AD, Helming L, Schiebe S, Wegener I, Hafner M, Beales M, Kontgen F, Lengeling A (2004) The phosphatidylserine receptor has essential functions during embryogenesis but not in apoptotic cell removal. J Biol 3(4):15PubMedCrossRefGoogle Scholar
  15. 15.
    Kunisaki Y, Masuko S, Noda M, Inayoshi A, Sanui T, Harada M, Sasazuki T, Fukui Y (2004) Defective fetal liver erythropoiesis and T lymphopoiesis in mice lacking the phosphatidylserine receptor. Blood 103(9):3362–3364PubMedCrossRefGoogle Scholar
  16. 16.
    Li MO, Sarkisian MR, Mehal WZ, Rakic P, Flavell RA (2003) Phosphatidylserine receptor is required for clearance of apoptotic cells. Science 302(5650):1560–1563PubMedCrossRefGoogle Scholar
  17. 17.
    Schneider JE, Bose J, Bamforth SD, Gruber AD, Broadbent C, Clarke K, Neubauer S, Lengeling A, Bhattacharya S (2004) Identification of cardiac malformations in mice lacking Ptdsr using a novel high-throughput magnetic resonance imaging technique. BMC Dev Biol 4(1):16PubMedCrossRefGoogle Scholar
  18. 18.
    Manaka J, Kuraishi T, Shiratsuchi A, Nakai Y, Higashida H, Henson P, Nakanishi Y (2004) Draper-mediated and phosphatidylserine-independent phagocytosis of apoptotic cells by Drosophila hemocytes/macrophages. J Biol Chem 279(46):48466–48476PubMedCrossRefGoogle Scholar
  19. 19.
    Hong JR, Lin GH, Lin CJ, Wang WP, Lee CC, Lin TL, Wu JL (2004) Phosphatidylserine receptor is required for the engulfment of dead apoptotic cells and for normal embryonic development in zebrafish. Development 131(21):5417–5427PubMedCrossRefGoogle Scholar
  20. 20.
    Williamson P, Schlegel RA (2004) Hide and seek: the secret identity of the phosphatidylserine receptor. J Biol 3(4):14PubMedCrossRefGoogle Scholar
  21. 21.
    Beninati S, Piacentini M (2004) The transglutaminase family: an overview: minireview article. Amino Acids 26(4):367–372PubMedGoogle Scholar
  22. 22.
    Falasca L, Iadevaia V, Ciccosanti F, Melino G, Serafino A, Piacentini M (2005) Transglutaminase type II is a key element in the regulation of the anti-inflammatory response elicited by apoptotic cell engulfment. J Immunol, 174(11):7330–7340PubMedGoogle Scholar
  23. 23.
    Szondy Z, Sarang Z, Molnar P, Nemeth T, Piacentini M, Mastroberardino PG, Falasca L, Aeschlimann D, Kovacs J, Kiss I, Szegezdi E, Lakos G, Rajnavolgyi E, Birckbichler PJ, Melino G, Fesus L (2003) Transglutaminase 2-/- mice reveal a phagocytosis-associated crosstalk between macrophages and apoptotic cells. Proc Natl Acad Sci USA 100(13):7812–7817PubMedCrossRefGoogle Scholar
  24. 24.
    Sakai K, Busby WH Jr, Clarke JB, Clemmons DR (2001) Tissue transglutaminase facilitates the polymerization of insulin-like growth factor-binding protein-1 (IGFBP-1) and leads to loss of IGFBP-1’s ability to inhibit insulin-like growth factor-I-stimulated protein synthesis. J Biol Chem 276(12):8740–8745PubMedCrossRefGoogle Scholar
  25. 25.
    Candi E, Paradisi A, Terrinoni A, Pietroni V, Oddi S, Cadot B, Jogini V, Meiyappan M, Clardy J, Finazzi-Agro A, Melino G (2004) Transglutaminase 5 is regulated by guanine-adenine nucleotides. Biochem J 381(Pt 1):313–319PubMedGoogle Scholar
  26. 26.
    Takeuchi T, Watanabe Y, Takano-Shimizu T, Kondo S (2006) Roles of jumonji and jumonji family genes in chromatin regulation and development. Dev Dyn 235(9):2449–2459PubMedCrossRefGoogle Scholar
  27. 27.
    Ayoub N, Noma K, Isaac S, Kahan T, Grewal SI, Cohen A (2003) A novel jmjC domain protein modulates heterochromatization in fission yeast. Mol Cell Biol 23(12):4356–4370PubMedCrossRefGoogle Scholar
  28. 28.
    Hewitson KS, McNeill LA, Riordan MV, Tian YM, Bullock AN, Welford RW, Elkins JM, Oldham NJ, Bhattacharya S, Gleadle JM, Ratcliffe PJ, Pugh CW, Schofield CJ (2002) Hypoxia-inducible factor (HIF) asparagine hydroxylase is identical to factor inhibiting HIF (FIH) and is related to the cupin structural family. J Biol Chem 277(29):26351–23655PubMedCrossRefGoogle Scholar
  29. 29.
    Mazure NM, Brahimi-Horn MC, Berta MA, Benizri E, Bilton RL, Dayan F, Ginouves A, Berra E, Pouyssegur J (2004) HIF-1: master and commander of the hypoxic world. A pharmacological approach to its regulation by siRNAs. Biochem Pharmacol 68(6):971–980PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Nitu Tibrewal
    • 1
  • Tong Liu
    • 2
  • Hong Li
    • 2
  • Raymond B. Birge
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyUMDNJ-New Jersey Medical SchoolNewarkUSA
  2. 2.Center for Advanced Proteomics Research and Department of Biochemistry and MolecularBiologyUMDNJ-New Jersey Medical School Cancer CenterNewarkUSA

Personalised recommendations