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- ItemSomente MetadadadosImmobilized Cratylia mollis lectin: An affinity matrix to purify a soybean (Glycine max) seed protein with in vitro platelet antiaggregation and anticoagulant activities(Elsevier B.V., 2011-01-01) Silva, Mariana C. C. [UNIFESP]; Santana, Lucimeire A. [UNIFESP]; Silva-Lucca, Rosemeire A.; Lima, Amanda L. R.; Ferreira, Joana G. [UNIFESP]; Paiva, Patricia M. G.; Coelho, Luana C. B. B.; Oliva, Maria L. V. [UNIFESP]; Zingali, Russolina B.; Correia, Maria T. S.; Universidade Federal de Pernambuco (UFPE); Universidade Federal de São Paulo (UNIFESP); Univ Estadual Oeste Parana; Universidade Federal do Rio de Janeiro (UFRJ)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.