Blood Clotting in Invertebrates

  • S. Srimal
Part of the Progress in Molecular and Subcellular Biology book series (PMSB, volume 15)


Well before clotting evolved, innate mechanisms served to prevent loss of body fluid from an injured site. Injury to an animal can result in a loss of body fluid and favour attack and entry of opportunistic pathogens. Irrespective of the level of organization, each organism is equipped with defense mechanisms to counteract this threat.


Horseshoe Crab Spiny Lobster Invertebr Pathol Octo Nate Sialic Acid Binding Lectin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Aketagawa J, Miyata T, Ohtsubo S, Nakamura T, Hayashida H, Miyata T, Iwanaga S (1986) Primary structure of Limulus anticoagulant anti-lipopolysaccharide factor. J Biol Chem 261: 7375–7365Google Scholar
  2. Armstrong PB (1977) Interaction of the motile blood cells of the horseshoe crab, Limuluspolyphemus. Studies on contact paralysis of pseudopodial activity and cellular overlapping in vitro. Exp Cell Res 107: 127–138PubMedCrossRefGoogle Scholar
  3. Armstrong PB (1979) Motility of the Limulus blood cell. J Cell Sci 37: 169–180PubMedGoogle Scholar
  4. Armstrong PB (1985) Adhesion and motility of the blood cells of Limulus. In: Cohen WD (ed) Blood cells of marine invertebrates: experimental systems in cell biology and comparative physiology. Alan R Liss, New YorkGoogle Scholar
  5. Armstrong PB (1991) Cellular and humoral immunity in the horseshoe crab. In: Gupta AP (ed) Limulus polyphemus immunology of insects and other arthropods. CRC Press, Boca Raton, pp 3–17Google Scholar
  6. Armstrong PB, Quigley JP (1991) α2-macroglobulin: a recently discovered defense system in arthropods. In: Gupta AP (ed) Immunology of insects and other arthropods. CRC Press, Boca RatonGoogle Scholar
  7. Armstrong PB, Rickles FR (1982) Endotoxin-induced degranulation of the Limulus amebocyte. Exp Cell Res 140: 15–24PubMedCrossRefGoogle Scholar
  8. Armstrong PB, Quigley JP, Rickles FR (1990) TheLimulus blood cell secretes α2-macroglobulin when activated. Biol Bull 178: 137–143CrossRefGoogle Scholar
  9. Bang FB (1956) A bacterial disease of Limulus polyphemus. Bull Johns Hopkins 98: 325–350Google Scholar
  10. Bang FB (1967) Serological responses among invertebrates other than insects. Fed Proc 26:1680–1684PubMedGoogle Scholar
  11. Barker WH Jr, Bang FB (1966) The effect of infection by Gram-negative bacteria and their endotoxins, on the blood clotting mechanisms of the crustacean Sacculina carcini, a parasite of the crab Carcinus Maenas. J Invertebr Pathol 8: 88–97PubMedCrossRefGoogle Scholar
  12. Bishayee S, Dorai DT (1980) Isolation and characterisation of a sialic binding lectin (carcinoscorpin) from Indian horseshoe crab C. rotundicauda. Biochem Biophys Acta 623: 89–97PubMedGoogle Scholar
  13. Bohn H (1986) Hemolymph clotting in insects. In: Brehelin M (ed) Immunity in invertebrates. Springer, Berlin Heidelberg New York, pp 188–207CrossRefGoogle Scholar
  14. Boman HG, Hultmark D (1987) Cell free immunity in insects. Annu Rev Microbiol 41: 103–126PubMedCrossRefGoogle Scholar
  15. Brandin ER, Pistole TG (1983) Polyphemin: a techoic acid-binding lectin from the horseshoe crab Limulus polyphemus. Biochem Biophys Res Commun 113: 611–617PubMedCrossRefGoogle Scholar
  16. Cannon GW, Tsuchiya M, Rittschof D, Bonaventura J (1986) Magnesium dependence of endotoxin-induced degranulation of Limulus amebocytes. Biol Bull 143: 548–567Google Scholar
  17. Campbell-Wilkes L (1973) PhD Dissertation, Northwestern Univ, Ann Arbor, MI, univ microfilms 73–30 763Google Scholar
  18. Cheng SM, Suzuki A, Zon G, Liu TY (1986) Characterisation of a complementary deoxyribonucleic acid for the coagulogen of Limulus polyphemus. Biochem Biophys Acta 868: 1–8PubMedGoogle Scholar
  19. Cornick JW, Stewart JE (1968) Interaction of the pathogen Gaffkya homari with natural defense mechanism ofHomarus americanus. J Fish Res Board 25: 695–709CrossRefGoogle Scholar
  20. Davie EW, Fujikawa K, Kisiel W (1991) the coagulation cascade: initiation, maintenance and regulation. Biochemistry 30: 10363–10370PubMedCrossRefGoogle Scholar
  21. Doolittle RF, Riley M (1990) The amino-terminal sequence of lobster fibrinogen reveals common ancestry with vitellogenins. Biochem Biophys Res Commun 167: 16–19PubMedCrossRefGoogle Scholar
  22. Dona van MA, Laue TM (1991) A novel trypsin inhibitor from the hemolymph of the horseshoe crab Limulus polyphemus. J Biol Chem 266: 2121–2125Google Scholar
  23. Dorai DT, Srimal S, Somasundaran M, Bachhawat BK, Balganesh TS (1982a) Recognition of 2-keto-deoxyoctonate in bacterial cell wall and lipopolysaecharides by the sialic acid binding lectin from the horseshoe crab. C. rotundicauda. Biochem Biophys Res Commun 104: 141–147PubMedCrossRefGoogle Scholar
  24. Dorai DT, Srimal S, Somasundaran M, Bachhawat BK, Balganesh TS (1982b) Oft the multispecifity of carcinoscorpin, the sialic acid binding lectin from the horseshoe crab. C. rotundicauda. FEBS Lett 148: 98–102CrossRefGoogle Scholar
  25. Dumont JN, Anderson E, Winner G (1966) Some cytologic characteristics of the hemocytes of Limulus during clotting. J Morphol 119: 181–208PubMedCrossRefGoogle Scholar
  26. Durliat M (1985) Clotting processes in crustacea decapoda. Biol Rev 60: 473–603CrossRefGoogle Scholar
  27. Durliat M, Vranckx R (1983) Analysis of clotting defects in diseased lobsters 2. Proteins of diseased lobster hemolymph. Comp Biochem Physiol 76A 103–108CrossRefGoogle Scholar
  28. Durliat M, Vranckx R (1989) Relationships between plasma and hemocyte proteins in decapoda. Comp Biochem Physiol 92B: 595–603Google Scholar
  29. Fine structure, morphological changes during coagulation and localization of clotting factors and antimicrobial substances. Cell tissue Res 266: 137–147Google Scholar
  30. Florkin M (1962) Comparative Biochemistry IV, a comprehensive treatise. Florkin M, Mason HS (eds) Academic Press, London, pp 435–482Google Scholar
  31. Fuji N, Minetti CAS A, Nakhasi H, Chen S, Barbehenn E, Nunes PH, Nguyen NY (1992) Isolation, cDNA cloning and characterisation of an 18 kDa hemagglutinin and amebocyte aggregation factor from Limulus polyphemus. J Biol Chem 267: 22452–22459Google Scholar
  32. Fuller GM (1978) Are lobster fibrinogen and cold in soluble-immunoglobulin related molecules? Ann NY Acad Sci 312: 31–37PubMedCrossRefGoogle Scholar
  33. Fuller GM, Doolittle RF (1971a) Studies of invertebrate fibrinogen I. Purification and characterisation of fibrinogen from spiny lobster. Biochemistry 10: 1305–1310PubMedCrossRefGoogle Scholar
  34. Fuller GM, Doolittle RF (1971b) Studies of invertebrate fibrinogen. II. Transformation of lobster fibrinogen into fibrin. Biochemistry 10: 1311–1315PubMedCrossRefGoogle Scholar
  35. Halliburton WD (1885) On the blood of decapod crustacea. J Physiol 6: 300–335PubMedGoogle Scholar
  36. Hardy WB (1892) The blood corpuscles of the crustacea together with a suggestion as to the origin of the crustacean fibrin ferment. J Physiol 13: 24–28Google Scholar
  37. Hoess A, Watson S, Siber GR, Liddington R (1993) Crystal structure of an endotoxin-neutralizung protein from the horseshoe crab, Limulus anti LPS-factor, at 1.5 A resolution. EMBO J 12: 3351–3356PubMedGoogle Scholar
  38. Hose JE, Martin GG, Nguyen VA, Rosenstein T (1987) Cytochemical features of shrimp hemocytes. Biol Bull 173: 176–185CrossRefGoogle Scholar
  39. Hose JE, Martin GG, Gerard AS (1990) A decapod hemocyte classification scheme integrating morphology, cytochemistry, and function. Biol Bull 178: 33–45CrossRefGoogle Scholar
  40. Iwanaga S (1993) TheLimulus clotting reaction. Curr Opin Immunol 5: 74–82PubMedCrossRefGoogle Scholar
  41. Iwanaga S, Morita T, Miyata T, Nakamura T, Aketagawa J (1986) The hemolymph coagulation system in invertebrate animals. J Protein Chem 5: 225–268CrossRefGoogle Scholar
  42. Iwanaga S, Miyata T, Tokunaga F, Muta T (1992) Molecular mechanism of hemolymph clotting system inLimulus. Thromb Res 68: 1–32PubMedCrossRefGoogle Scholar
  43. Kenney DM, Belamarich FA, Shepro D (1972) Aggregation of horseshoe crab(Limulus polyphemus) amebocytes and reversible inhibition of aggregation of EDTA. Biol Bull 143: 548–567PubMedCrossRefGoogle Scholar
  44. Kopacek P, Hall M, Soderhall K (1993) Characterisation of a clotting protein isolated from plasma of the freshwater crayfishPacifastacus leniusculus. Eur J Biochem 213: 591–597PubMedCrossRefGoogle Scholar
  45. Lewis T (1979) In: Cohen E (ed) Biomedical applications of the horseshoe crab (Limulidae). Alan R Liss, New York, p 682Google Scholar
  46. Levin J (1967) Blood coagulation and endotoxin in invertebrates. Fed Proc 26: 1707–1712Google Scholar
  47. Levin J, Bang FB (1964a) The role of endotoxin in the extracellular coagulation of Limulus blood. Bull John Hopkins 115: 265–274Google Scholar
  48. Levin J, Bang FB (1964b) A description of cellular coagulation in Limulus. Bull John Hopkins 115: 337–345Google Scholar
  49. Liu T, Lin Y, Cislo T, Mineti CAS A, Baba JMK, Liu TY (1991) Limunectin: A phosphochholine-binding protein from Limulus amebocytes with adhesion-promoting properties. J Biol Chem 266: 14813–14821PubMedGoogle Scholar
  50. Loeb L (1902) On the blood lymph cells and inflammatory processes of Limulus. J Med Res 7: 145–158PubMedGoogle Scholar
  51. Loeb L (1920) The movements of the amebocytes and the experimental production of amebocyte (cellfibrin) tissue. Wash Univ Stud 8:3Google Scholar
  52. Lorand L (1972) Fibrinoligase: the fibrin-stabilising factor system of blood plasmas. Ann NY Acad Sci 202: 1–348Google Scholar
  53. Lorand L, Conrad SM (1984) Transglutaminases. Mol Cell Biochem 58: 9–35PubMedCrossRefGoogle Scholar
  54. Madaras F, Chew MY, Parkin JD (1981) Purification and characterisation of the sand crab (Ovalipes bipustulatus) coagulogen (fibrinogen). Thromb Haemostasis 45: 77–81Google Scholar
  55. Marchalonis J J, Edelman GM (1968) Isolation and characterisation of a hemagglutinin from L.ployphemus. J Mol Biol 32: 265–275CrossRefGoogle Scholar
  56. Martin GG, Hose JE, Kim JJ (1987) Structure of hematopoietic nodules in the ridgeback prawn, Sicyonia ingentis: light and electron microscopic observations. J Morphol 192: 193–204CrossRefGoogle Scholar
  57. Martin GG, Hose JE, Omori S, Chong C, Hoodboy T, McKrell N (1991) Localization and roles of coagulogen and transglutaminase in hemolymph coagulation in decapod crustaceans. Comp Biochem Physiol 100B: 517–522Google Scholar
  58. Minetti CAS A, Lin Y, Cislo T, Liu TY (1991) Purification and characterization of an endotoxin-binding protein with protease inhibitory activity from amebocytes of Limulus polyphemus. J Biol Chem 266: 20773–20780PubMedGoogle Scholar
  59. Miyata T, Hiranaga M, Umezu M, Iwanaga S (1984) Amino acid sequence of the coagulogen from Limulus polyphemus hemocytes. J Biol Chem 259: 8924–8933PubMedGoogle Scholar
  60. Miyata T, Matsumoto H, Hattori M, Sakaki Y, Iwanaga S (1986) Two types of coagulogen mRNAs found in horseshoe crab (Tachypleus tridentatus) hemocytes: Molecular cloning and nuclotide sequences. J Biochem (Tokyo) 100: 213–220Google Scholar
  61. Morita T, Tanaka S, Nakamura T, Iwanaga S (1981) D-glucan mediated coagulation pathway found inLimulus amebocytes. FEBS Lett 129: 318–321CrossRefGoogle Scholar
  62. Muta T, Miyata T, Tokunaga F, Nakamura T, Iwanaga S (1987) Primary structure of anti-lipopolysaccharide factor from American horseshoe crab, Limulus polyphemus. J Biochem (Tokyo) 101: 1321–1330Google Scholar
  63. Muta T, Miyata T, Misumi Y, Tokunaga F, Nakamura T, Toh Y, Ikehara Y, Iwanaga S (1991a) Limulus factor C: An endotoxin sensitive serine protease zymogen with a mosaic structure of complement-like, epidermal growth factor-like, and lectin-like domains. J Biol Chem 266: 6554–6561PubMedGoogle Scholar
  64. Muta T, Hashimoto R, Oda T, Miyata T, Iwanaga S (1991b) Limulus clotting enzyme and factor B associated with endotoxin-sensitive coagulation cascade: Novel serine protease zymogens with a new type of “disulfide-knotted domain”. Throm Haemostasis Abstr 65: 935Google Scholar
  65. Nakamura S, Iwanaga S, Harada T, Niwa M (1976a) A clottable protein (coagulogen) from amebocyte lysate of Japanese horseshoe crab (Tachypleus tridentatus). J Biochem (Tokyo) 80: 1011–1021Google Scholar
  66. Nakamura S, Iwanaga S, Harada T, Niwa M (1976b) A clottable protein (coagulogen) of horseshoe crab hemocytes: structural change of its polypeptide chain during gel formation. J Biochem (Tokyo) 80: 649–652Google Scholar
  67. Nakamura T, Morita T, Harada-Suziki T, Iwanaga S (1985) Intracellular proclotting enzyme in (Tachypleus tridentatus) hemocytes: Its purification and properties. J Biochem (Tokyo) 97: 1561–1574Google Scholar
  68. Nakamura S, Horuchi T, Morita T, Iwanaga S (1986) Purification and properties of intracellular clotting factor, factor B from horseshoe crab(Tachypleus tridentatus) hemocytes. J Biochem (Tokyo) 99: 847–857Google Scholar
  69. Nakamura T, Hirai T, Tokunaga F, Kawabata S, Iwanaga S (1987) Purification and amino acid sequence of Kunitz-type protease inhibitor found in hemocytes of the horseshoe crab (Tachypleus tridentatus). J Biochem (Tokyo) 101: 1297–1306Google Scholar
  70. Nakamura T, Tokunaga F, Morita T, Iwanaga S (1988a) Intracellular serine-protease zymogen, factor C from horseshoe crab hemocytes: its activation by synthetic lipid A analogues and acidic phospholipids. Eur J Biochem 176: 89–94PubMedCrossRefGoogle Scholar
  71. Nakamura T, Furunaka H, Miyata T, Tokunaga F, Muta T, Iwanaga S, Niwa M, Takao T, Shimonishi Y (1988b) Tachyplesin, a class of antimicrobial peptide from hemocytes of the horseshoe crab(Tachypleus tridentatus). J Biol Chem 263: 16709–16713PubMedGoogle Scholar
  72. Needham AE (1970) Haemostatic mechanisms in the invertebrata. Symp Zool Sqc Lond 27: 19–44Google Scholar
  73. Ornberg RL, Reese TS (1981) Beginning of exocytosis captured by rapid-freezing of Limulus amebocyte. J Cell Biol 90: 40–54PubMedCrossRefGoogle Scholar
  74. Quigley JP, Armstrong PB(1983) An endopeptidase inhibitor similar to a?-macroglobulin, detected in the hemolymph of an invertebrate Limulus polyphemus. J Biol Chem 258: 7903–7906PubMedGoogle Scholar
  75. Ravindranath MH (1980) Hemocytes in hemolymph coagulation of arthropods. Biol Rev 55:139–170CrossRefGoogle Scholar
  76. Shishikura F, Sekiguchi K (1983) Agglutinins in the horseshoe crab hemolymph: Purification of a potent agglutinin of horse erythrocytes from the hemolymph of Tachypleus tridentatus, the japanese horseshoe crab. J Biochem 93: 1539–1546Google Scholar
  77. Söderhäll K (1981) Fungal cell wall ß-1-3 glucans induce clotting and phenol-oxidase attachment to foreign surfaces of crayfish hemocyte lysate. Dev Comp Immunol 5: 565–573PubMedGoogle Scholar
  78. Söderhäll K (1982) Prophenol-oxidase activating system and melanization - a recognition mechanism of arthropods? A review Dev Comp Physiol 6: 601–611Google Scholar
  79. Söderhäll K, Cerenius L (1992) Crustacean immunity. Annu Rev Fish Dis 1:3–23CrossRefGoogle Scholar
  80. Söderhäll K, Hall L (1984) LPS induced activation of prophenol-oxidase activating system in crayfish hemocyte lysate. Biochem Biophys Acta 797: 99–104CrossRefGoogle Scholar
  81. Söderhäll K, Levin J, Armstrong PB (1985) The effect of ß-l-3 glucans on the blood coagulation and amebocyte release in the horseshoe crab Limulus polyphemus. Biol Bull 169: 661–674CrossRefGoogle Scholar
  82. Spurling NW (1981) Comparative physiology of blood clotting. Comp Biochem Physiol 68A: 541–548CrossRefGoogle Scholar
  83. Srimal S (1984) The role of lipopolysaccharide binding proteins in the coagulation system ofCarci-noscorpious rotundicauda. PhD Thesis, Univ Calcutta, IndiaGoogle Scholar
  84. Srimal S, Bachhawat B (1985) Blood Brother: a living fossil helps save human lives. 51–55 Science Age Nov-Dec: 51–55Google Scholar
  85. Srimal S, Miyata T, Kawabata S, Iwanaga S (1985) the complete amino acid sequence of coagulogen isolated from the south-east Asian horseshoe crab Carcinoscorpious rotundicauda. J Biochem (Tokyo) 98: 305–318Google Scholar
  86. Srimal S, Dorai DT, Somasundaran M, Bachhawat BK, Miyata T (1985) A new hemagglutinin from the amebocytes of the horseshoe crab Carcinoscorpious rotundicauda. Purification and role in cell aggregation. Biochem J 230: 321–327PubMedGoogle Scholar
  87. Tait J (1911) Types of crustacean blood coagulation. J Mar Biol Assoc (UK) 9: 191–198CrossRefGoogle Scholar
  88. Toh Y, Mizutani A, Tokunaga F, Muta T, Iwanaga S (1991) Structure of hemocytes of the Japanese horseshoe crab Tachypleus tridentatus Google Scholar
  89. Tokunaga F, Miyata T, Nakamura T, Morita T, Kuma K, Miyata T, Iwanaga S (1987) Lipopolysaccharides-sensitive serine protease zymogen (Factor C) of horseshoe crab hemocytes: identification and alignment of proteolytic fragment produced during the activation show that it is a novel type of serine protease. Eur J Biochem 167: 405–416PubMedCrossRefGoogle Scholar
  90. Tokunaga F, Nakajima H, Iwanaga S (1991) Purification and characterisation of lipopolysaccharides-sesnsitive serine protease zymogen (factor C) isolated from Limulus polyphemus hemocytes: a newly identified intracellular zymogen activated by a-Chymotrypsin, not by trypsin, J Biochem (Tokyo) 109: 150–157Google Scholar
  91. Tokunaga F, Yamada M, Miyata T, Ding YL, Hiranaga-Kawabata M, Muta T, Iwanaga S, Ichinose A, Davie EW (1993a) Limulus hemocyte transglutaminase: its purification, characterisation and identification of the intracellular substrates. J Biol Chem 268: 252–261PubMedGoogle Scholar
  92. Tokunaga F, Muta T, Iwanaga S, Ichinose A, Davie EW, Kuma K, Miyata T (1993b) Limulus hemocyte transglutaminase: cDNA cloning, amino acid sequence and tissue localization. J Biol Chem 268: 262–265PubMedGoogle Scholar
  93. Yaith P, Uhlenbruck G, Holz G (1979) Anti-glucoronyl activityLimulus Polyphemus agglutinin. Protides Biol Fluids Coll 27: 455–458Google Scholar
  94. Valembios P, Roch PH, Lassegues M (1988) Evidence of plasma clotting in earthworm. J Invertebr Pathol 51: 221–228CrossRefGoogle Scholar
  95. Wainwright NR, Miller RJ, Paus E, Novitsky TJ, Fletcher MA, McKenna TM, Williams T (1990) In: Nowotny A, Spitzer JJ, Ziegler EJ (eds) Cellular and molecular aspects of endotoxin reactions. Elsevier New York, pp 315–325Google Scholar
  96. White M, Wainwright N, Novitsky TJ (1995) Isolation of three lectin from the horseshoe rab Limulus polyphemus. (in prep)Google Scholar
  97. Wilson J, Rickles FR, Armstrong PB, Lorand L (1992) Ne (glutamyl) lysine crosslinks in the blood clot of the horseshoe crab Limulus polyphemus. Biophys Biochem Res Commun 188: 655–661CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1996

Authors and Affiliations

  • S. Srimal
    • 1
  1. 1.Molecular Biophysics UnitIndian Institute of ScienceBangaloreIndia

Personalised recommendations