Partial purification and characterization of a lectin from Crotalaria retusa L. seeds
DOI:
https://doi.org/10.22481/exon.v10i2.20468Keywords:
Crotalaria retusa L, hemagglutinating activity, lectin, FaboideaeAbstract
Lectins are proteins or glycoproteins of non-immune origin, which bind reversibly to specific carbohydrates and are widely distributed in nature. This work aimed the partial purification and characterization of a lectin from Crotalaria retusa L. seeds (CrL). The crude extract was obtained by crushing the seeds and subsequent homogenization in 0.15M NaCl solution. The lectin was able to agglutinate all human erythrocytes tested, with a minimum concentration of 7.81μg/ml. CrL was inhibited by L-fucose, N-acetylgalactosamine and D-galactose in the minimum concentrations of 12.5mM, 25mM and 25mM, respectively, but it was not dependent on divalent cations. CrL was partially purified by size exclusion chromatography on Sephadex G-75 and exhibited a molecular mass of 30 kDa in polyacrylamide gel electrophoresis (SDS-PAGE). The lectin was stable at 100oC for 30 minutes and the hemagglutinating activity was maintained at pH 7.0 – 9.0.
Downloads
References
Van Damme, E.J.M.; Peumans, W.J.; Barre, A.; Rouge, P. Plant lectins: A composite of several distinct families of structurally and evolutionary related proteins with diverse biological roles, Crit. Rev. Plant Sci., 17 (1998) 575–692.
Sharon, N.; Lis, H. History of lectins: from hemagglutinins to biological recognition molecules, Glycobiology, 14 (2004) 53R-62R.
Gabius, H.J. Glycans: bioactive signals decoded by lectins, Biochem. Soc. Trans, 36 (2008) 1491-1496.
Santos, A.F.S.; da Silva, M.D.C.; Napoleão, T. H.; Paiva, P.M.G.; Correia, M.T.S.; Coelho , L.C.B.B. Lectins: Function, structure, biological properties and potential applications, Current Topics in Peptide & Protein Research, 15 (2014) 41-62.
Charungchitrak, S.; Petsom, A.; Sangvanich, P. Antifungal and antibacterial activities of lectin from the seeds of Archidendron jiringa Nielsen, Food Chem, 126 (2011) 1025 – 1032.
Santos, A.F.S.; Napoleão, T.H.; Bezerra, R.F.; Carvalho, E.V.M.M.; Correia, M. T. S.; Paiva, P.M.G.; Coelho, L.C.B.B. Strategies to obtain lectins from distintic sources, in: Berhardt, L.V. (Ed.), Advances in Medicine and Biology, v. 63, Nova Science Publishers, Inc., 2013, pp. 33-60.
Kaur, M.; Singh, K.; Rup, P.J.; Saxena, A.K.; Khan, R.H.; Ashraf, M.T.; Kamboj, S.S.; Singh, J. Atuber lectin from Arisaema helleborifolium Schott with anti-insect activity against melon fruit fly, Bactrocera cucurbitae (Coquillett) and anti-cancer effect on Insecticide Activity of Lectins and Secondary Metabolites 593 human cancer cell., 445 (2006) Arch. Biochem. Biophys. 156-165.
Souza, J.D.; Silva, M.B.R.; Argolo, A.C.C.; Napoleão, T.H.; Sá, R.A.; Correia, M.T.S.; Paiva, P.M.G.; Silva, M.D.C. & Coelho, L.C.B.B. A new Bauhinia monandra galactose-specific lectin purified in milligram quantities from secondary roots with antifungal and termiticidal activities. Int. Biodeterior. Biodegradation, 65 (2011b) 696-702.
Costa, R.M.P.B.; Vaz, A.F.M.; Oliva, M.L.V.; Coelho, L.C.B.B.; Correia, M.T.S.; Carneiro-da- Cunha, M.G. 2010. A new mistletoe Phthirusa pyrifolia leaf lectin with antimicrobial properties. Process Biochem, 45:526-533.
Wang, H.; Ng, T.B. A lectin with some unique characteristics from the samta tomato, Plant Physiol. Bioch, 44 (2006) 181-185.
Rüdiger, H. Plant lectins – more than just tools for glycoscientists: occurrence, structure and possible functions of plant lectins, Acta Anat, 161 (1998) 130-152.
Macedo, M.L.R; Oliveira, C.F.R; Oliveira, C.T. Insecticidal Activity of Plant Lectins and Potential Application in Crop Protection, Molecules, 20 (2015) 2014-2033.
Peumans, W.J.; Van Damme, E.J.M. Lectins as plant defense proteins, Rev. Plant Physiol, 109 (1995) 347-352.
Panda, P.K.; Mukhopadhyay, S.; Behera, B.; Bhol, C.S.; Dey, S.; Das, D.N.; Sinha, N.; Bissoyi, A.; Pramanik, K.; Maiti, T.K.; Bhutia, S.K. Antitumor effect of soybean lectin mediated through reactive oxygen species-dependent pathway, Life Sci, 111 (2014) 27–35.
Maciel, E.V.; , Araújo-Filho, V.S.; Nakazawa, M.; Gomes, Y.M.; Coelho, L.C.; Correia, M.T. Mitogenic activity of Cratylia mollis lectin on human lymphocytes, Biologicals, 32 (2004) 57-60.
Silva, M.C.C.; Santana, L.A.; Mentele, R.; Ferreira, R.S.; Miranda, A.; Silva-Lucca, R.A. Purification, primary structure and potential functions of a novel lectin from Bauhinia forficata seeds, Process Biochem, 7 (2012) 41049-1059.
Araújo, R.M.S; Vaz, A.F.M.; Aguiar, J.S.; Coelho, L.C.B.B; Paiva, P.M.G; Melo, A.M.M.; Silva, T.G.; Correia, M.T.S. Lectin from Crataeva tapia bark exerts antitumor, antiinflammtory and analgesic activities. Nat. prod. bioprospect, 1 (2011) 97–100.
Cummings, R.D. Lectins as tools for glycoconjugate purification and characterization. In Glycosciences: Status and Perspectives; Gabius, H.-J., Gabius, S., Eds.; Chapman & Hall: London, UK, 1997; pp. 191–199.
Lam, S.K., Ng, T.B. Lectins: production and practical applications, Appl. Microbiol. Biotechnol, 89 (2011) 45–55.
Kennedy, J.F., Paiva, P.M.G., Corella, M.T.S., Cavalcanti, M.S.M., Coelho, L.C.B.B. Lectins, versatile proteins of recognition: a review, Carbohydr. Polym, 26 (1995) 219-230.
Lewis, G.P., Schrire, B., Mackinder, B., Lock, M. Legumes of the world. Royal Botanical Garden: Kew; 2005.
Flores, A.S., Miotto, S.T.S. Aspectos fitogeográficos das espécies de Crotalaria L. (Leguminosae, Faboideae) na Região Sul do Brasil, Acta Bot. Brasilica, 19 (2005) 245-249.
Devendra, B.N., Srinivas, N., Solmon, K.S. A comparative pharmacological and phytochemical analysis of in vivo & in vitro propagated Crotalaria species, Asian Pac. J. Trop. Med, 5 (2012) 37-41.
Honório Júnior, J.E.R., Soares, P.M., Melo, C.L., Arruda Filho, A.C.V., Sena Filho, J.G., Barbosa Filho, J.M., Sousa, F.C.F.Atividade farmacológica da monocrotalina isolada de plantas do gênero Crotalaria, Ver. Bras. Farmacogn, 20 (2010) 453-458.
Ersson, B., Aspberg, K., Porath, J. The Phytohemagglutinin from sunn hemp seeds (Crotalaria juncea). Purification by biospecific affinity chromatography, Biochim. Biophys. Acta, 310 (1973) 446-452.
Ersson, B. A phytohemagglutinin from sunn hemp seeds (Crotalaria juncea). II Purification by a high capacity biospecific adsorbent and its physicochemical properties, Biochim. Biophys. Acta, 494 (1977) 51-60.
Khang, N.Q.; Jean-Luc, G.; Johan, H. A blood group A specific lectin from the seeds of Crotalaria striata, Biochim. Biophys. Acta, 1033 (1990) 210-213.
Rego, E.J.L.; De Carvalho, D.D.; Marangoni, S.; De Oliveira, B.; Novello, J.C. Lectins from seeds of Crotalaria pallida (smooth rattlebox), Phytochemistry, 60 (2002) 441-446.
Pando, L.A..; De Carvalho, D.D.; Toyama, M.H.; Di Ciero, L.; Novello, J.C.; Pascholatti, S.F. ;Marangoni, S. Purification and characterization of a lectin from Crotalaria paulina seeds, Protein J, 23 (2004) 437-444.
Kaur, N.; Singh, J.; Kamboj, S.S. Affinity purification and characterization of a seed lectin from Crotalaria medicaginea, Indian J. Biochem. Biophys, 39 (2002) 49-54.
Bradford, M.M. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of dye binding, Anal. Biochem, 72 (1976) 248-254.
Alsever J.B; Ainslie R.B. A new method for the preparation of dilute blood plasma and the operation of a complete transfusion service, N.Y. State J. Med; 41 (1941) 126-131.
Laemmli, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 227 (1970) 680-685.
Howard, J.A.; Edward, A. Z. J.; Khsushi, L.M. A Comparison of the Binding Specificities of Lectins from Ulex Europaeus and Lotus Tetragonolobus, Immunological Communications, 6 (1977) 585-602.
Pando, S.C.; Macedo, M.L.R.; Freire, M.G.M.; Toyama, M.H.; Novello, J.C.; Marangoni, S. Biochemical characterization of a lectina from Delonix regia seeds, J. Protein Chem, 21 (2002) 279-285.
Silva, H.C.; Bari, A.U.; Pereira-Junior, F.N.; Simões, R.C.; Barroso- Neto, I.L.; Nobre, C.B.; Pereira, M.G.; Nascimento, K.S.; Rocha, B.A.; Delatore, P.; Nagano, C.S.; Assreuy, A.M.; Cavada, B.S. Purification and partial characterization of a new pro-inflammatory lectin from Bauhinia bauhinioides Mart (Caesalpinoideae) seeds, Protein Pept. Lett, 18 (2011) 396-402.
Loris, R. Principles of structures of animal and plant lectins, Biochim. Biophys. Acta,, 1572 (2002) 198-208.
Carvalho, A.S.; Silva, M.V.; Gomes, F.S.; Paiva, P.M.G.; Malafaia, C.B.; Silva, T.D. Purification, characterization and antibacterial potential of a lectin isolated from Apuleia leiocarpa seeds, Int. J. Biol. Macromol, 75 (2015)402-408.
Oliveira, M.D.L.; Andrade, C.A.S.; Santos-Magalhães, N.S.; Coelho, L.C.B.B.; Teixeira, J.A. ;Carneiro-da-Cunha, M.G.;Correia, M.T.S. Purification of a lectin from Eugenia uniflora L. seeds and its potential antibacterial activity, Lett. Appl. Microbiol, 46 (2008) 371-376.
Silva, H.C.; Nagano, C.S.; Souza, L.A.G.; Nascimento, K.S.; Isidro, R.; Delatore, P. Purification and primary structure determination of a galactose-specific lectin from Vatairea guianensis Aublet seeds that exhibits vasorelaxant effect, Process Biochem, 47 (2012) 2347-2355.
Silva, M.; Silva, S.; Nascimento, K.; Nagano, C.S.; Cavada, B.S. Physico-chemical characterization and partial sequence of a lectin from Canavalia bonariensis Lindl seeds, BMC Proc 8 (Suppl 4) (2014) 227.
Osterne, V.J.S.; Santiago, M.Q.; Pinto Junior, V.R.; Cajazeiras, J.B.; Correia, J.L.A.; Leitão, C.C.F. Purification, partial characterization, and CNBr-Sepharose immobilization of a vasorelaxant glucose/mannose lectin from Canavalia virosa seeds, Appl. Biochem. Biotechnol, 172 (2014) 3342-3353.
Sharma, A.; Wong, J.H.; Lin, P.;Chan, Y.S.; Ng, T.B. Purification and characterization of a lectin from Indian cultivar of French bean seeds, Protein Pept. Lett, 17 (2010) 221-227.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2019 Exatas Online
This work is licensed under a Creative Commons Attribution 4.0 International License.
Você é livre para:
Compartilhar - copia e redistribui o material em qualquer meio ou formato; Adapte - remixe, transforme e construa a partir do material para qualquer propósito, mesmo comercialmente. Esta licença é aceitável para Obras Culturais Livres. O licenciante não pode revogar essas liberdades, desde que você siga os termos da licença.
Sob os seguintes termos:
Atribuição - você deve dar o crédito apropriado, fornecer um link para a licença e indicar se alguma alteração foi feita. Você pode fazer isso de qualquer maneira razoável, mas não de uma forma que sugira que você ou seu uso seja aprovado pelo licenciante.
Não há restrições adicionais - Você não pode aplicar termos legais ou medidas tecnológicas que restrinjam legalmente outros para fazer qualquer uso permitido pela licença.
