Abstract
Hemocytes of the horseshoe crab contain a family of arthropod peptide antibiotics, termed the tachyplesin family, and an antibacterial protein, called anti-LPS factor, of which the former is located in the small (S) granules and the latter in the large (L) granules of the hemocytes (1–5). In our ongoing studies on granular components, we have identified a novel defensin-like substance, named big defensin, present in both L- and S-granules (6). This substance strongly inhibits the growth of Gram-negative and Gram-positive bacteria, and fungi, such as Candida albicans (Table 1). The isolated big defensin consists of a total of 79 amino acid residues, in which the COOH-terminal region composed of 37 amino acids resembles mammalian defensins. Big defensin, however, is distinct from the mammalian defensins in molecular size, the latter of which have 29–34 amino acid residues in common (7,8). The disulfide motif in the defensin-like domain of big defensin is identical to that of β-defensin from bovine neutrophils but not to that of classical defensins (9,10). Furthermore, the structural organization of big defensin differs markedly from those of insect defensins not only in disulfide bridge locations but also in the molecular size (11,12).
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References
Toh, Y., Mizutani, A., Tokunaga, F., Muta, T., and Iwanaga, S. (1991) Morphology of the granular hemocytes of the Japanese horseshoe crab Tachypleus tridentatus and immunocytochemical localization of clotting factors and antimicrobial substances. Cell Tissue Res. 266, 137–147.
Iwanaga, S., Muta, T., Shigenaga, T., Seki, N., Kawano, K., Katsu, T., and Kawabata, S. (1994) Structure-function relationships of tachyplesins and their analogues, in Ciba Foundation Symposium 186, Antimicrobial Peptides (Boman, H. G., Marsh, J., and Goode, J. A., eds.), Wiley, Chichester, England, pp. 160–175.
Iwanaga, S., Muta, T., Shigenaga, T., Miura, Y., Seki, N., Saito, T., and Kawabata, S. (1994) Role of hemocyte-derived granular components in invertebrate defense. Ann. NY Acad. Sci. 712, 102–116.
Muta, T. and Iwanaga, S. (1996) Invertebrate immunology, in Progress in Molecular and Subcellular Biology 15 (Rinkevich, B. and Muller, W. E. G., eds.), Springer-Verlag, Berlin, pp. 154–189.
Kawabata, S., Muta, T., and Iwanaga, S. (1996) Clotting cascade and defense molecules found in hemolymph of horseshoe crab, in New Directions in Invertebrate Immunology (Söderhäll, K., Iwanaga, S., and Vasta, G. R., eds.), SOS Publications, Fair Haven, NJ, pp. 255–284.
Saito, T., Kawabata, S., Shigenaga, T., Cho, J., Nakajima, H., Hirata, M., and Iwanaga, S. (1995) A novel big defensin identified in horseshoe crab hemocytes: isolation, amino acid sequence and antibacterial activity. J. Biochem. (Tokyo), 117, 1131–1137.
Lehrer, R. I., Ganz, T., and Selsted, M. E. (1991) Defensins endogenous antibiotic peptides of animal cells. Cell 64, 229, 230.
Selsted, M. E. and Harwig, S. S. L. (1989) Determination of the disulfide array in the human defensin HNP-2. J. Biol. Chem. 264, 4003–4007.
Selsted, M. E., Tang, Y.-Q., Morris, W. L., McGuire, P. A., Novotny, M. J., Smith, W., Henschen, A. H., and Cullor, J. S. (1993) Purification, primary structure, and antibacterial activities of β-defensins, a new family of antimicrobial peptides from bovine neutrophils. J. Biol. Chem. 268, 6641–6648.
Tang, Y.-Q. and Selsted, M. E. (1993) Characterization of the disulfide motif in BNBD-12, an antimicrobial β-defensin peptide from bovine neutrophils. J. Biol. Chem. 268, 6649–6653.
Lambert, J. Keppi, E., Dimarcq, J-L., Wicker, C., Reichhart, J-M., Dunbar, B., Lepage, P., Dorsselaer, A.V., Hoffmann, J., Fothergill, J., and Hoffmann, D. (1989) Insect immunity: isolation from immune blood of the dipteran Phormia terranovae of two insect antibacterial peptides with sequence homology to rabbit lung macrophage bactericidal peptides. Proc. Natl. Acad. Sci. USA 86, 262–266.
Kuzuhara, T., Nakajima, Y., Matsuyama, K., and Natori, S. (1990) Determination of the disulfide array in sapecin, an antibacterial peptide of Sarcophaga peregrina (flesh fly). J. Biochem. (Tokyo) 107, 514–518.
Saito, T., Kawabata, S., Hirata, M., and Iwanaga, S. (1995) A novel type of limulus lectin-L6 Purification, primary structure, and antibacterial activity. J. Biol. Chem. 270, 14,493–14,499.
Okino, N., Kawabata, S., Saito, T., Hirata, M., Takagi, T., and Iwanaga, S. (1995) Purification, characterization, and cDNA cloning of a 27-kDa lectin (L10) from horseshoe crab hemocytes. J. Biol. Chem. 270, 31,008–31,015.
Nowak, T. P. and Barondes, S. H. (1975) Agglutinin from Limulus polyphemus: purification with formalinized horse erythrocytes as the affinity adsorbent. Biochim. Biophys. Acta 393, 115–123.
Bishayee, S. and Dorai, D. T. (1980) Isolation and characterization of a sialic acid-binding lectin (carcinoscorpin) from Indian horseshoe crab Carcinoscorpius rotundicauda. Biochim. Biophys. Acta 623, 89–97.
Tsuboi, I., Matsukawa, M., Sato, N., and Kimura, S. (1993) Isolation and characterization of a sialic acid-specific binding lectin from the hemolymph of Asian horseshoe crab, Tachypleus tridentatus. Biochim. Biophys. Acta 1156, 255–262.
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Kawabata, Si., Iwanaga, S. (1997). Big Defensin and Tachylectins-1 and -2. In: Shafer, W.M. (eds) Antibacterial Peptide Protocols. Methods In Molecular Biology™, vol 78. Humana Press. https://doi.org/10.1385/0-89603-408-9:51
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DOI: https://doi.org/10.1385/0-89603-408-9:51
Publisher Name: Humana Press
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