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A novel mannose-binding lectin from plasma of Labeo rohita

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Abstract

A new mannose-binding lectin was purified from plasma of freshwater fish, rohu (Labeo rohita) by ammonium sulphate precipitation followed by DEAE-cellulose ion-exchange chromatography. The hemagglutinating activity is strong for neuraminidase-treated rabbit erythrocytes. Mannose, glucose and their derivatives inhibit hemagglutination. The apparent molecular weight was determined to be 210 kDa in native PAGE. Analysed on SDS-PAGE under reducing and non-reducing conditions, the protein migrated as a single band of mol.wt. 40,000. The lectin is acidic in nature showing a pI of 4.5. It is a glycoprotein containing complex glycan part as indicated by carbohydrate analysis using high-pressure anion exchange chromatography with a pulsed amperometric detector. The N-terminal sequence (WLNGIGTQIPMKITT) shows no significant homology with known proteins. It appears to be a C-type lectin as Ca2+ is essential for carbohydrate-binding activity. Methyl -α- D-mannopyranoside was found to be the most potent inhibitor among the monosaccharides and disaccharides tested. Purified lectin was found to induce intracellular superoxide production following opsonization of E. coli by its own macrophages.

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REFERENCES

  • Das, H.R., Jayaraman, V. and Bhattacharya, I. 1999.Carbohydrate analysis of bradyrhizobial (NC 92) lipopolysaccharides by high performance-anion exchange chromatography with pulsed amperometric detection.Biosc. Rep.19:219–225.

    Article  CAS  Google Scholar 

  • Davis, B.J. 1964. Disc electrophoresis-II Method and application to human proteins.Ann. NY Acad. Sci.121:404–427.

    PubMed  CAS  Google Scholar 

  • Drickamer, K. 1999.C-type lectin-like domains.Curr. Opin. Struc. Biol. 9:585–590.

    Article  CAS  Google Scholar 

  • Ewart, K.V. Johnson, S.C. and Ross, N. W. 1999.Identification of pathogen binding lectin in salmon serum.Comp. Biochem. & Physiol. Part C. 123:9–15.

    CAS  Google Scholar 

  • Fock, W.L, Chen, C.L., Lam, T.J and Sin, Y.M. 2000. Isolation and characterization of a serum lectin from blue gourami, Trichogaster trichopterus.Fish Shellfish Immunol.10:489–504.

    Article  PubMed  CAS  Google Scholar 

  • Fock, W.L., Chen, C.L., Lam, T.J and Sin, Y.M. 2001. Roles of an endogenous serum lectin in the immune protection of blue gourami, Trichogaster trichopterus (Pallus) against Aeromonas hydrophila.Fish Shellfish Immunol.11:101–113.

    Article  PubMed  CAS  Google Scholar 

  • Gallagher, J.T. 1984.Carbohydrate-binding properties of lectins: A possible approach to lectin nomenclature and classification. Biosci. Rep.4:621–632.

    Article  PubMed  CAS  Google Scholar 

  • Gercken, J. and Renwrantz, L. 1994.A new mannan-binding lectin from serum of eel (Anguilla anguilla L). Isolation, characterization and comparison with fucose specific serum lectin.Comp. Biochem. Physiol. Biochem. Mol. Biol.108:449–461.

    Article  PubMed  CAS  Google Scholar 

  • Hardy, M.R. Townsend, R.R. and Lee, Y.C. 1988. Monosaccharide analysis of glycoconjugates by anion-exchange chromatography with pulsed amperometric detection.Anal. Biochem.170:54–62.

    Article  PubMed  CAS  Google Scholar 

  • Hosono, M. Kawauchi, H. Nitta, K. Takayanagi, Y. Shiokawa, H. Mineki, R. and Murayama, K. 1993a. Three rhamnose-binding lectins from Osmerus eperlanus mordax (olive rainbow smelt) roe.Biol. Pharm. Bull. 16: 239–243.

    PubMed  CAS  Google Scholar 

  • Hosono, M. Kawauchi, H. Nitta, K. Takayanagi, Y. Shiokawa, H. Mineki, R. and Murayama, K.1993b.Purification and characterization of Silurus asotus (catfish) roe lectin.Biol. Pharm. Bull16: 1–5.

    PubMed  CAS  Google Scholar 

  • Iwasaki, M., Seko, A., Kitajima, K., Inoue, Y. and Inoue, S.1992. Fish egg glycophosphoproteins have species specific N-linked glycan units previously found in a storage pool of free glycan chains.J. Biol. Chem.267:24287–24296.

    PubMed  CAS  Google Scholar 

  • Jensen, L.E. Thiel, S. Petersen, T. E. and Jensenius, J.C. 1997. A rainbow trout lectin with multimeric structure.Comp. Biochem. Physiol.116B:385–390.

    CAS  Google Scholar 

  • Laemmli, U.K. 1970.Cleavage of structural proteins during assembly of head of bacteriophage T4. Nature.227:680–685.

    Article  PubMed  CAS  Google Scholar 

  • Lowry, O.M. Rosenbrough, N.J., Fars, A.L. and Randall, R.J. 1951. Protein measurement with Folin phenol reagent.J. Biol. Chem. 193:265–275.

    PubMed  CAS  Google Scholar 

  • Lu, J. 1997. Collectins: collectors of microorganisms for the innate immune system.BioEssays.19:509–518.

    Article  PubMed  CAS  Google Scholar 

  • Manihar, S.R. 1992. Studies on fresh water fish lectins. Ph.D. Thesis, Delhi University.

  • Manihar, S. R. and Das, H. R. 1990. Isolation and characterization of new lectin from plasma of fish Channa punctatus.Biochim. Biophys. Acta. 1036:162–165.

    PubMed  CAS  Google Scholar 

  • Manihar, S. R. Das, R.H. and Das, H. R. 1991.A T-antigen binding lectin from Channa leucopunctatus (Murrel) plasma. Carbohydrate Research.213: 251–261.

    Article  PubMed  CAS  Google Scholar 

  • Markert, M. Andrews, P.C. and Babior, B.M. (1984). Measurements of O- 2production by human neutrophils. The preparation and assay of NADPH oxidase-containing particles from human neutrophils.Methods Enymol.105:358–365.

    Article  CAS  Google Scholar 

  • Muramoto, K. Kamiya, H. 1992. The amino-acid sequence of a lectin from conger myriaster, skin mucus.Biochim. Biophys. Acta.1116:129–136.

    PubMed  CAS  Google Scholar 

  • Papac, D.I., Briggs, J.B. Chin, E.T. and Jones, A.J.S. 1998. A high-throughput microscale method to release N-linked oligosaccharides from glycoproteins for matrix-assisted laser desorption/ ionization time of flight mass spectrometric analysis.Glycobiol. 8:445–454.

    Article  CAS  Google Scholar 

  • Ofek, I. and Sharon, N. 1988. Lectinophagocytosis: A molecular mechanism of recognition between cell surface sugars and lectins in the phagocytosis of bacteria.Infect. Immun.56:539–547.

    PubMed  CAS  Google Scholar 

  • Ottinger, C.A. Johnson, S.C. Ewart, K.V. Brown, L.L. and Ross, N.W. 1999. Enhancement of anti-Aeromonas salmonicida activity in Atlantic salmon (Salmo salar) macrophages by a mannosebinding lectin. Comp. Biochem. Physiol. Part C123:53–59.

    Article  CAS  Google Scholar 

  • Roberton, E.F., Dannelly, H.K., Malloy, P.J. and Reeves, H.C. 1987. Rapid Isoelectric focusing in a vertical polyacrylamide minigel system.Anal. Biochem.167:290–294.

    Article  Google Scholar 

  • Tateno, H. Saneyoshi, A. Ogawa, T. Muramoto, K. Kamiya, H. and Saneyoshi, M. 1998. Isolation and characterization of rhamnosebinding lectins from eggs of steelhead trout homologous to low density lipoprotein receptor superfamily.J.Biol.Chem.273: 19190–19197.

    Article  PubMed  CAS  Google Scholar 

  • Vasta, G.R., Quesenberry, M., Ahmed, H. and O'Leary, N. 1999. C-type lectins and galectins mediate innate and adaptive immune functions: their roles in the complement activation pathway.Dev. Comp. Immunol.23:401–420.

    Article  PubMed  CAS  Google Scholar 

  • Weis, W.I. and Drickamer, K. (1994). Trimeric structure of C-type mannose-binding protein.Structure2:1227–1240.

    Article  PubMed  CAS  Google Scholar 

  • White C. A. and Kennedy J.F. 1986. Oligosaccharides. In: Carbohydrate analysis, a practical approach. pp. 37–54. Edited by Chaplin and Kennedy. IRL Press, Oxford.

    Google Scholar 

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Authors and Affiliations

  1. Lectin Research Laboratory, Centre for Biochemical Technology, Delhi University Campus, Mall Road, Delhi, 110007, India

    S. Mitra & H.R. Das

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  1. S. Mitra
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  2. H.R. Das
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Mitra, S., Das, H. A novel mannose-binding lectin from plasma of Labeo rohita . Fish Physiology and Biochemistry 25, 121–129 (2001). https://doi.org/10.1023/A:1020545510295

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