Immobilized Cratylia mollis lectin: An affinity matrix to purify a soybean (Glycine max) seed protein with in vitro platelet antiaggregation and anticoagulant activities

Immobilized Cratylia mollis lectin: An affinity matrix to purify a soybean (Glycine max) seed protein with in vitro platelet antiaggregation and anticoagulant activities

Author Silva, Mariana C. C. Autor UNIFESP Google Scholar
Santana, Lucimeire A. Autor UNIFESP Google Scholar
Silva-Lucca, Rosemeire A. Google Scholar
Lima, Amanda L. R. Google Scholar
Ferreira, Joana G. Autor UNIFESP Google Scholar
Paiva, Patricia M. G. Google Scholar
Coelho, Luana C. B. B. Google Scholar
Oliva, Maria L. V. Autor UNIFESP Google Scholar
Zingali, Russolina B. Google Scholar
Correia, Maria T. S. Google Scholar
Institution Universidade Federal de Pernambuco (UFPE)
Universidade Federal de São Paulo (UNIFESP)
Univ Estadual Oeste Parana
Universidade Federal do Rio de Janeiro (UFRJ)
Abstract Lectins, proteins which recognize selectively carbohydrates, have been used as affinity chromatography to purify glycoproteins. Cratylia mollis seed lectin (Cramoll 1,4), of mannose/glucose binding class, immobilized on Sepharose CL-4B, was used as affinity matrix. This paper describes the purification by Cramoll 1,4-Sepharose matrix, and characterization of an anti-platelet and anticoagulant soybean (Glycine max) protein, ApcSP, and its in vitro platelet antiaggregation and anticoagulant activities. SDS-PAGE of ApcSP purified under reducing condition revealed a single glycosilated band of 51 kDa. the N-terminal 10-residue sequence of ApcSP is EGQFGPMIQS, distinct to other soybean seed proteins, such as peroxidase, lectin, 2S albumin and trypsin inhibitor. Deconvolution of CD spectrum indicated 35% alpha-helix, 17% beta-strand, 22% turn and 26% unordered structure; ApcSP fluorescence spectrum showed a maximum emission around 339 nm. the hemostatic parameters revealed inhibition of collagen (p<0.001), thrombin (p<0.05) and ADP (p<0.001)-induced platelet aggregation in the presence of ApcSP (2 mu M), in relation to positive control. the soy protein prolonged the blood coagulation time (activated partial thromboplastin time, more affected, and prothrombin time). the results indicated that immobilized Cramoll 1,4 lectin has the potential to isolate soybean glycoproteins and ApcSP may be important for anti-thrombotic and anticoagulant therapy. (C) 2010 Elsevier B.V. All rights reserved.
Keywords Antiaggregant
Anticoagulant
Cramoll 1,4-Sepharose
Glycine max
Glycoprotein
Lectin
Language English
Sponsor Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundacao de Amparo a Ciencia e Tecnologia do Estado de Pernambuco (FACEPE)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Date 2011-01-01
Published in Process Biochemistry. Oxford: Elsevier B.V., v. 46, n. 1, p. 74-80, 2011.
ISSN 1359-5113 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 74-80
Origin http://dx.doi.org/10.1016/j.procbio.2010.07.017
Access rights Closed access
Type Article
Web of Science ID WOS:000286538300009
URI http://repositorio.unifesp.br/handle/11600/33287

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