Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Purification and Characterization of Hemagglutinating Protein from Rhizome of Alisma orientale

택사(Alismatis Rhizoma) Hemagglutinating Protein의 정제와 특성

  • Published : 1995.08.01
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Abstract

Lectin was purified by using $(NH_4)_2SO_4$, DEAE-cellulose ion-exchange chromatography and Sephadex G-150 column chromatography from Alismatis Rhizoma(AR). The specific activity of AR lectin was 50, 441units/mg, and purification folds were 114. The AR lectin agglutinated human erythrocytes of all types(A, B, O, AB). The molecular weight of AR lectin was estimated about 90, 500 daltons by gel filtration and each subunits were 42,000, 27,000 and 22,500 daltons on SDS-PAGE respectively. The hemagglutinating activity of the lectin was inhibited by sialic acid, glucose, ribose, galactose, sucrose, and lactose. It was also inhibited by cations such as $Hg^{++},\;Fe^{++},\;Cu^{++}\;and\;Pb^{++}$.

택사로부터 황산암모늄 분별 침전, DEAE-cellulose ion exchange chromatography, Sephadex G-150 chromatography 등의 방법을 이용하여 렉틴을 분리, 정제하였다. 정제한 렉틴의 분자량은 gel filtration법에 의해 측정한 결과 90,500 dalton이었으며, SDS polyacrylamide gel 전기영동을 실시한 결과 subunit 분자량은 각각 42,000, 27,000, 22,500 dalton으로 나타나므로 택사 렉틴이 heterotrimer임을 알았다. 정제된 택사 렉틴은 사람 적혈구의 경우 모든 혈액형에 대하여 응집현상을 나타내었으며 돼지, 쥐 및 개 등의 적혈구에 대해서도 모두 응집현상이 나타나 혈구 비특이성임을 보여주었다. 또한 이 렉틴은 sialic acid, glucose, ribose, sucrose, lactose, galactose 등 당에 의해서, 그리고 $Hg^{2+},\;Fe^{2+},\;Cu^{2+}$ 이온 등에 의해 적혈구 응집력이 저해되었다.

Keywords

References

  1. Science v.177 Cell-Agglutinating and sugar-specific proteins Sharon,N.;Lis,H.
  2. Annu. Rev. Microbiol. v.35 Interaction of bacteria and fungi with lectins and lectin-like substances Pistole,N.
  3. Science v.246 Lectins as cell recognition molecules Sharon,N.;Lis,H.
  4. J. Immunology v.108 The characterization of purified erythrocyte agglutinins from two invertebrate species Finstad,C.D.;Litman,G.W.;Finstad,J.;Good,R.A.
  5. Biochemistry v.22 Purification and properties of two hemagglutinins of the mushroom Laccaria amethystina Guillot,J.;Genaud,L.;Gueugnot,J.;Damez,M.
  6. Nature v.292 Defining a lectin Dixon,H.B.F.
  7. Cancer Research v.20 An initiator of mitosis cultures of normal human leukocytes Noweoo,P.C.
  8. Clin. Exp. Immunol. v.13 Reconstitution of immunocompetence in B cells by addition of concanavalin A or concanavalin A-treated thymus cells Sjoberg,O.
  9. J. Immunology v.148 Characterization of proteoglycan-reactive T cell lines and hybridomas from mice with proteoglycan-induced arthritis Leroux,J.Y.;Poole,A.R.;Webber,C.;Vipparti,V.;Choi,H.U.
  10. Immunology v.75 Pattern of lectin binding to murine T lymphocytes Sowalsky,R.A.;Fox,B.S.
  11. J. Biol. Chem. v.249 Wheat germ agglutinin Nagata,Y.;Burger,M.M.
  12. J. Clin. Microbiol. v.10 Lectins on diagnostic microbiology : Use of wheat germ agglutinin for laboratory idendification of Neisseria genorrhoeae Schaefer,R.L.;Keller,K.F.;Doyle,R.J.
  13. Biochemistry v.13 Characterization of two plant lectins from Ricinus communis and their quantitative interaction with a murine lymphoma Nicolson,G.L.;Blaustein,J.;Etzler,M.E.
  14. J. Biol. Chem. v.249 Purification and characterization of two major toxoc proteins from seeds of Abrus precatorius Wei,C.H.;Hartman,F.;Pfuderer,P.;Yang,W.K.
  15. Eur. J.Clin. Microbiol. v.3 Lectins in diagnostic microbiology Doyle,R.;Keller,K.
  16. Biochem. J. v.96 The gel filteration behavior of proteins related to their molecular weight over a wide range Andrews,P.
  17. J. Biol. Chem. v.244 The reliability of molecular weight determinations by dodecyl dulfate-poly-acrylamide gel electrophoresis Webber,K.;Osborn,M.
  18. Annal. Biochem. v.72 A rapid and sensitive method for the quantitation microgam quantities of protein utilizing of principle of protein-dye binding Bradford,M.M.
  19. Arch. Pharm. Res. v.3 Isolation, purification and characterization of phytohemagglutinating proteins from Korean natural products Chung,S.R.;Jeume-chung,K.H.;Kim,K.A.
  20. J. Biol. Chem. v.246 Studies on a phytohemagglutinin from the lentil Howard,I.K.;Sage,H.J.;Stein,M.D.
  21. J. Biol. Chem. v.249 Subunit structure of soybean agglutinin Lotan,R.;Siegelman,H.W.;Lis,H.;Sharon,N.
  22. J. Biol. Chem. v.257 Isolation and characterization of Viseumin, a toxic lectin from Viscum album L.(Mistretoe) Olsnens,S.;Stripe,F.;Dandvig,K.;Pihl,A.
  23. Biochemistry v.16 Purification and properties of the hemagglutinin from Maclura pomifera seeds Bausch,J.N.;Poretz,R.D.
  24. J. Biol. Chem. v.250 The covalent and three dimensional structure of concanavalin A Becker,J.W.;Reeke,G.N.Jr.;Wang,J.L.;Cunningham,B.A.;Edelman,G.M.