Abstract
Two types of lectins from Dioclea sericea seeds were obtained by saline extraction and precipitation with 30-70% s (NH4)2SO4 or by 2.5% trichloroacetic acid precipitation. The P4 lectin was purified by affinity chromatography on Sephacryl S- 200; the P2 lectin was purified by hydrophobic chromatography on Phenyl- Sepharose. The following properties were determined for each lectin: carbohydrate specificity, pI. subunit Mr, molecular weight of the native protein, carbohydrate content, minimal inhibitory capacity, animal erythrocyte agglutination, amino acid composition, and N-terminal sequence of the α., ß and ɣ chains of P4 lectin and α. chain of the P2 lectin. The lectin content in mature seeds was established.
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References
Ainouz I.I., Moreira, R. A., Campos, F.D.A.P., Richardson, M., Begbie, R., Stewart, J.C., Watt, W.B. & Pusztai A. 1987. "The isolation and amino acid sequence of the b- and g-subunits of the lectin from the seeds of Dioclea grandiflora. Phytochemistry "26., 1435-1440.
Andrews, P. 1965. "The gel filtration behaviour of proteins related to their molecular weights over a wide range" Biochem. J. 96, 595-606.
A.O.A.C. 1975, Official methods of analysis of the Association of Agricultural Chemists "12 Edition, Washington.
Bernal, H. & Jiménez, L. 1990. "Haba Criolla". SECAB. Programa de recursos vegetales del SECAB. Editorial Guadalupe. Bogotá, Colombia.
Bollag, D., Edelstein S. & Stuart., J. 1991 "Isoelectric focusing (IEF)". Protein Methods. Wiley-liss, New York.pp 162-164.
Bowles, D.J. & Papin, D.J. 1988. "Traffic and assembly of concanavalin A". Trends Biochem. Sci., 13, 60-64.
Deutscher, M. 1990. "Guide to protein purificación" Meth. Enzymol. 182, 339-402.
Dubois, M., Gilles, K., Hamilton, J., Rebers, P. & Smith, F. 1956. "Colorimetric method for determination of sugars and related substances". Anal. Chem., 28, 350-356.
Galvis, M. & Múnera, M. 1974. "Evaluación de la proteína de Canavalia ensiformis". Tesis, Departamento de Química, Universidad Nacional de Colombia, Bogotá.
Gupta, D., Osscarson, S., Raja, T., Stanley, P., Toone, E. & Brawer, C. 1996. "A comparison of the fine saccharide-binding specificity of Dioclea grandiflora lectin and concanavalin A. "Eur. J. Biochem.. 242, 320-326.
Hamerstrand, G.E., Black, L.T. & Glover, J.D. 1981. "Trypsin inhibitors in soy products: Modification of the standard analytical procedure" Cereal Chem, 58, 42-45.
Hirs, W. & Stein, H. 1956. "Peptides obtained by Chymotryptic hidrolysis of performic acid-oxidized ribonuclease. A partial structural formula for the oxidized protein". J. Biol. Chem. 221, 151-169.
Hoeffer. 1992. "2-D Gel Electrophoresis". Scientific Instruments. 136-140.
Humboldt, A. & Bonpland, A. 1823. "Nova genera et species plantarum". Sixieme partie botanique. Apaud Gide Filium, Bibliopolam. No. 20, 457-458, 576.
Laemmli, U. 1970. "Cleavage of structural proteins during the assembly of the head of Bacteriophage T4" Nature, 227, 680-685.
Lakhtin, V.M. 1989. "Biotechnology of lectins" Biotekhnologiya. 5, 676-91.
Lis, H. & Sharon, N. 1986. "Applications of lectins" in The lectins, (Liener I..E..Sharon N., Goldstein I.J., eds.) Academic Press, Orlando, pp 293-370.
Matsudaira, P. 1987. "Sequence from picomole quantities of proteins electroblotted onto polyvinylidene diflouride membranes". J. Biol. Chem., 262, 10035-10038.
Melgarejo, L. M. 1997. Localización intracelular de las lectinas P, y P, de Dioclea lehmanni y estudio de la homología de la región amino terminal de las lectinas P2 presentes en Dioclea grandiflora y Canavalia ensiformis. Tesis Doctorado. Depto Quimica, Universi- dad Nacional de Colombia.
Melgarejo, L.M. & Pérez, G. 1997. Immunolocalization of the lectins P, and P, from Dioclea lehmanni seeds". Plant Cell Physiol., 38, 480-483.
Moreira, R., Barros, A., Stewart, J. & Puzstai, A. 1983. "Isolation and characterization of a lectin from the seeds of Dioclea grandiflora." Planta, 158, 63-69.
Moreira, R., Monteiro, A., Horta, A., Oliveira, J. & Cavada, B. 1997. "Isolation and characterization of Dioclea altissima Var. Megacarpa seed lectin". Phytochemistry. 46. 139-144.
Pérez, G. 1998. Isolation and characterization of a novel lectin from Dioclea lehmanni (Fabaceae) seeds. Int.J.Biochem Cell Biol., 30, 843-853.
Pérez, G., Hernández, M. & Mora, E. 1989. "Isolation and characterization of a lectin from the seeds of Dioclea lehmanni ". Phytochemistry, 29, 1.749-1.759.
Pérez, G., Pérez, C., Sousa, A, B., Moreira, A, R. & Richardson, M. 1991. "Comparison of the amino acid sequences of the lectins from seeds of Dioclea lehmanni and Canavalia maritima. " Phytochemistry, 30, 2619-2621.
Sierra, A.Y. 1997. "Extracción, purificación y caracterización parcial de las lectinas presentes en las semillas de Dioclea sericea " Tesis, Departamento de Química, Universidad Nacional de Colombia, Bogotá.
Steyermark, B. 1961. Quantitative organic microanalysis..Second edition, Academic Press, New York, pp 188-209.
Villarraga M. & Peña, M. 1979. "Purificación y caracterización parcial de la lectina presente en la Erythrina rubrinervia". Tesis, Departamento de Química. Universidad Nacional de Colombia, Bogotá.
Weber, K. & Osborn, M. 1975. "Proteins and sodium dodecyl sulfate: Molecular weight determination of polyacrylamide gels and related procedures" en "The proteins". 3a edición, Neurath H., Hill R.L.. (eds.), Academic Press. New York Vol. 1. pp 179-223.
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