Download PDF
Thromb Haemost 2000; 84(06): 1087-1094
DOI: 10.1055/s-0037-1614175
Review Article
Schattauer GmbH

Occurrence of Amphoterin (HMG1) as an Endogenous Protein of Human Platelets that Is Exported to the Cell Surface upon Platelet Activation

Ari Rouhiainen
1   From Finnish Red Cross Blood Transfusion Service, Helsinki, Finland
2   Laboratory of Molecular Neurobiology, Institute of Biotechnology, Department of Biosciences, Division of Biochemistry, University of Helsinki, Finland
,
Shinji Imai
2   Laboratory of Molecular Neurobiology, Institute of Biotechnology, Department of Biosciences, Division of Biochemistry, University of Helsinki, Finland
3   Department of Anatomy, Shiga University of Medical Science, Shiga-ken, Japan
,
Heikki Rauvala
2   Laboratory of Molecular Neurobiology, Institute of Biotechnology, Department of Biosciences, Division of Biochemistry, University of Helsinki, Finland
,
Jaakko Parkkinen
1   From Finnish Red Cross Blood Transfusion Service, Helsinki, Finland
› Author AffiliationsThe authors thank Dr. Riitta Kekomäki for help in platelet binding studies. The excellent technical assistance of Anne Remes is gratefully acknowledged. This work was supported by the Sigrid Jusélius Foundation and the Academy of Finland.
Further Information

Publication History

Received 14 February 2000

Accepted after resubmission 14 July 2000

Publication Date:
13 December 2017 (online)

    Permissions and Reprints

Summary

Amphoterin (HMG1) is a 30-kD heparin-binding protein which is functionally associated with the outgrowth of cytoplasmic processes in developing neurones. Amphoterin has been shown to mediate adhesive and proteolytic interactions at the leading edge of motile cells. Recently it was shown that inhibition of amphoterin interactions with its cell surface receptor (RAGE) suppresses tumour growth and metastasis. In this work we have identified amphoterin polypeptide and its mRNA in human platelets. Amphoterin had a cytoplasmic localisation in resting platelets according to subcellular fractionation studies and immunogold electronmicroscopy. After platelet activation, part of amphoterin was associated with the external surface of plasma membrane. Externalisation of amphoterin during platelet activation was also detected in immunofluorescence studies. Amphoterin was detectable in human serum (0.2 ng/ml) but not in plasma. Resting platelets treated with PGI2 and forskolin bound to immobilised recombinant amphoterin independently of divalent cations. The binding induced a spicular morphology in platelets, and was effectively inhibited by heparin. Amphoterinbinding protein components on the platelet surface were not identified, but amphoterin bound to phosphatidylserine and sulfatide in lipid binding assays. Our results suggest that amphoterin is an endogenous protein in human platelets, which is exported to the cell surface during platelet activation. Interaction of amphoterin with the platelet surface may be mediated by sulfoglycolipids and phospholipids.

Key words

Amphoterin - heparin - HMG1 - platelet - phosphatidylserine
 

  • References

  • 1 Rauvala H, Pihlaskari R. Isolation and some characteristics of an adhesive factor of brain that enhances neurite outgrowth in central neurons. J Biol Chem 1987; 262: 16625-35.
    PubMedGoogle Scholar
  • 2 Rauvala H, Merenmies J, Pihlaskari R, Korkolainen M, Huhtala M, Panula P. The adhesive and neuritepromoting molecule p30: Analysis of the aminoterminal sequence and production of antipeptide antibodies that detect p30 at the surface of neuroblastoma cells and of brain neurons. J Cell Biol 1988; 107: 2293-305.
    CrossrefPubMedGoogle Scholar
  • 3 Merenmies J, Pihlaskari R, Laitinen J, Wartiovaara J, Rauvala H. 30-kDa heparin binding protein of brain (amphoterin) involved in neurite out-growth. J Biol Chem 1991; 266: 16722-9.
    PubMedGoogle Scholar
  • 4 Parkkinen J, Raulo E, Merenmies J, Nolo R, Kajander EO, Baumann M, Rauvala H. Amphoterin, the 30-kDa protein in a family of HMG1-type polypeptides. J Biol Chem 1993; 268: 19726-38.
    PubMedGoogle Scholar
  • 5 Punnonen EL, Fages C, Wartiovaara J, Rauvala H. Ultrastructural localization of β-actin and amphoterin mRNA in cultured cells: Application of tyramide signal amplification and comparison of detection methods. J Histochemistry and Cytochemistry 1998; 47: 99-112.
    PubMedGoogle Scholar
  • 6 Fages C, Nolo R, Huttunen HJ, Eskelinen E, Rauvala H. Regulation of cell migration by amphoterin. J Cell Sci 2000; 113: 611-20.
    PubMedGoogle Scholar
  • 7 Hori O, Brett J, Slattery T, Cao R, Zhang J, Xian JChen, Nagashima M, Lundh ER, Vijay S, Nitecki D, Morser J, Stern D, Schmidt AM. The Receptor for Advanced Glycation End Products (RAGE) Is a Cellular Binding Site for Amphoterin. J Biol Chem 1995; 270: 25752-61.
    CrossrefPubMedGoogle Scholar
  • 8 Huttunen HJ, Fages C, Rauvala H. Receptor for advanced glycation end products (RAGE)-mediated neurite outgrowth and activation of NF-kappaB require the cytoplasmic domain of the receptor but different downstream signaling pathways. J Biol Chem 1999; 274: 19919-24.
    CrossrefPubMedGoogle Scholar
  • 9 Taguchi A, Blood DC, Del Toro G, Canet A, Lee DC, Qu W, Tanji N, Lu Y, Lalla E, Fu C, Hofmann MA, Kislinger T, Ingram M, Lu A, Tanaka H, Hori O’Ogawa S, Stern DM, Schmidt AM. Blockade of RAGE–amphoterin signalling suppresses tumour growth and metastases. Nature 2000; 405: 354-60.
    CrossrefPubMedGoogle Scholar
  • 10 Grosschedl R, Giese K, Pagel J. HMG domain proteins: architectural elements in the assembly of nucleoprotein structures. Trends Genet 1994; 10: 94-100.
    CrossrefPubMedGoogle Scholar
  • 11 Bustin M, Lehn DA, Landsman D. Structural features of the HMG chromosomal proteins and their genes. Biochim Biophys Acta 1990; 1049: 231-43.
    CrossrefPubMedGoogle Scholar
  • 12 Calogero S, Grassi F, Aguzzi A, Voigtlander T, Ferrier P, Ferrari S, Bianchi ME. The lack of chromosomal protein Hmg1 does not disrupt cell growth but causes lethal hypoglycaemia in newborn mice. Nat Genet 1999; 22: 276-80.
    CrossrefPubMedGoogle Scholar
  • 13 Merenmies J, Pihlaskari R, Laitinen J, Wartiovaara J, Rauvala H. 30-kDa heparin binding protein of brain (amphoterin) involved in neurite outgrowth. J Biol Chem 1991; 266: 16722-9.
    PubMedGoogle Scholar
  • 14 Nair SM, Zhao Z, Chou DKH, Tobet SA, Jungalwala FB. Expression of HNK-1 carbohydrate and its binding protein SBP-1, in apposing cell surfaces in cerebral cortex and cerebellum. Neuroscience 1998; 85: 759-71.
    CrossrefPubMedGoogle Scholar
  • 15 Wang H, Bloom O, Zhang M, Vishnubhakat JM, Ombrellino M, Che J, Frazier A, Yang H, Ivanova S, Borovikova L, Manogue KR, Faist E, Abraham E, Andersson J, Andersson U, Molina PE, Abumrad NN, Sama A, Tracey KJ. HMG-1 as a late mediator of endotoxin lethality in mice. Science 1999; 285: 248-51.
    CrossrefPubMedGoogle Scholar
  • 16 Ombrellino M, Wnag H, Ajemian MS, Talhouk A, Scher LA, Friedman SG, Tracey KJ. Increased serum concentrations of high-mobility group protein 1 in haemorrhagic shock. Lancet 1999; 354: 1446-7.
    CrossrefPubMedGoogle Scholar
  • 17 Parkkinen J, Rauvala H. Interactions of plasminogen and tissue plasminogen activator (t-PA) with amphoterin. J Biol Chem 1991; 266: 16730-5.
    PubMedGoogle Scholar
  • 18 Nordenman B, Bjork I. Purification of thrombin by affinity chromatography on immobilized heparin. Thromb Res 1977; 11: 799-808.
    CrossrefPubMedGoogle Scholar
  • 19 Thomas JO, Kornberg RD. An octamer of histones in chromatin and free in solution. Proc Natl Acad Sci 1975; 72: 2626-30.
    CrossrefPubMedGoogle Scholar
  • 20 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5.
    CrossrefPubMedGoogle Scholar
  • 21 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-9.
    PubMedGoogle Scholar
  • 22 Fox JEB, Reynolds CC, Boyles JK. Studying the platelet cytoskeleton in Trition X-100 lysates. Methods in Enzymology 1992; 215: 42-58.
    PubMedGoogle Scholar
  • 23 Bevers EM, Wiedmer T, Comfurius P, Shattil SJ, Weiss HJ, Zwaal RFA, Sims PJ. Defective Ca(2+)-induced microvesiculation and deficient expression of procoagulant activity in erythrocytes from a patient with a bleeding disorder: a study of the red blood cells of Scott syndrome. Blood 1992; 79: 380-8.
    PubMedGoogle Scholar
  • 24 Bellavite P, Andrioli G, Guzzo P, Arigliano P, Chirumbolo S, Manzato F, Santonastaso C. A colorimetric method for the measurement of platelet adhesion in microtiter plates. Anal Biochem 1994; 216: 444-50.
    CrossrefPubMedGoogle Scholar
  • 25 Folch J, Lees M, Stanley HS. A simple method for the isolation and purification of totel lipides from animal tissues. J Biol Chem 1957; 226: 497-509.
    PubMedGoogle Scholar
  • 26 Mohan PS, Laitinen J, Merenmies J, Rauvala H, Jungalwala FB. Sulfoglycolipids bind to adhesive protein amphoterin (P30) in the nervous system. Biochem Biophys Res Commun 1992; 182: 389-96.
    PubMedGoogle Scholar
  • 27 Broekman MJ. Homogenization by nitrogen cavitation technique applied to platelet subcellular fractionation. Methods in Enzymology 1992; 215: 21-32.
    PubMedGoogle Scholar
  • 28 Gogstad GO, Hagen I, Korsmo R, Solum NO. Characterization of the proteins of isolated human platelet alpha-granules. Evidence for a separate alpha-granule-pool of the glycoproteins IIb and IIIa. Biochim Biophys Acta 1981; 670: 150-62.
    CrossrefPubMedGoogle Scholar
  • 29 Fox JEB. The platelet cytoskeleton. Thromb Haemost 1993; 70: 884-93.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Savage B, Shattil SJ, Ruggeri ZM. Modulation of Platelet Function through Adhesion Receptors. J Biol Chem 1992; 267: 11300-6.
    PubMedGoogle Scholar
  • 31 Melloni E, Sparatore B, Patrone M, Pessino A, Passalacqua M, Pontremoli S. Identity in molecular structure between “differentiation enhancing factor” of murine erythroleukemia cells and the 30-kD heparin-binding protein of developing rat brain. Biochem Biophys Res Commun 1995; 210: 82-9.
    CrossrefPubMedGoogle Scholar
  • 32 Muesch A, Hartmann E, Rohde K, Rubartelli A, Sitia R, Rapoport TA. A novel pathway for secretory proteins?. Trends Biochem Sci 1990; 15: 86-8.
    CrossrefPubMedGoogle Scholar
  • 33 Hawrylowicz CM, Santoro SA, Platt FM, Unanue ER. Activated platelets express IL-1 acitivity. J Immunol 1989; 143: 4015-8.
    CrossrefPubMedGoogle Scholar
  • 34 Sedlmayr P, Blaschitz A, Wilders-Truschnig M, Tiran A, Dohr G. Platelets contain interleukin-1 alpha and beta which are detectable on the cell surface after activation. Scand J Immunol 1995; 42: 209-14.
    CrossrefPubMedGoogle Scholar
  • 35 Sawicki G, Sanders EJ, Salas E, Wozniak M, Rodrigo J, Radomski MW. Localization and translocation of MMP-2 during aggregation of human platelets. Thromb Haemost 1998; 80: 836-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 36 Sawicki G, Salas E, Murat J, Miszta-Lane H, Radomski MW. Release of gelatinase A during platelet activation mediates aggregation. Nature 1997; 386: 616-9.
    CrossrefPubMedGoogle Scholar
  • 37 Nair SM, Jungalwala FB. Characterization of a sulfoglucuronyl carbohydrate binding protein in the developing nervous system. J Neurochem 1997; 68: 1286-97.
    PubMedGoogle Scholar
  • 38 Roberts DD, Haverstick DM, Dixit VM, Frazier WA, Santoro SA, Ginsburd V. The platelet glycoprotein thrombospondin binds specifically to sulfated glycolipids. J Biol Chem 1985; 260: 9405-11.
    PubMedGoogle Scholar
  • 39 Uesugi H, Ozaki S, Sobajima J, Osakada F, Shirakawa H, Yoshida M, Nakao K. Prevalence and characterization of novel pANCA, antibodies to the high mobility group non-histone chromosomal proteins HMG1 and HMG2, in systemic rheumatic diseases. J Rheumatol 1998; 25: 703-9.
    PubMedGoogle Scholar
  • 40 Bustin M, Reisch J, Einck L, Klippel JH. Autoantibodies to nucleosomal proteins: antibodies to HMG-17 in autoimmune diseases. Science 1982; 4537: 1245-7.
    PubMedGoogle Scholar