Partial characterization and immobilization in CNBr-activated Sepharose of a native lectin from Platypodium elegans seeds (PELa) and comparative study of edematogenic effect with the recombinant form

被引：15

作者：

Araripe, David Alencar ^{[1
]}

Pinto-Junior, Vanir Reis ^{[1
]}

Bastos Neco, Antonio Hadson ^{[1
]}

Santiago, Mayara Queiroz ^{[1
]}

Silva Osterne, Vinicius Jose ^{[1
]}

Pires, Alana Freitas ^{[2
]}

Lossio, Claudia Figueiredo ^{[1
]}

Queiroz Martins, Maria Gleiciane ^{[1
]}

Almeida Correia, Jorge Luiz ^{[1
]}

Benevides, Raquel Guimaraes ^{[3
]}

Leal, Rodrigo Bainy ^{[4
]}

Sampaio Assreuy, Ana Maria ^{[2
]}

Nascimento, Kyria Santiago ^{[1
]}

Cavada, Benildo Sousa ^{[1
]}

机构：

[1] Univ Fed Ceara, Fortaleza, Ceara, Brazil

[2] Univ Estadual Ceara UECE, Fortaleza, Ceara, Brazil

[3] Univ Estadual Feira de Santana UEFS, Feira De Santana, BA, Brazil

[4] Univ Fed Santa Catarina, Florianopolis, SC, Brazil

来源：

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2017年 / 102卷

关键词：

PELa; Immobilization; Inflammation; VATAIREA-MACROCARPA SEEDS; PEANUT ARACHIS-HYPOGAEA; BINDING PLANT-LECTINS; RAT PAW EDEMA; NITRIC-OXIDE; ANTIBACTERIAL ACTIVITY; NEUTROPHIL MIGRATION; N-GLYCANS; PURIFICATION; PROTEINS;

D O I：

10.1016/j.ijbiomac.2017.03.193

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The lectin from Platypodium elegans seeds (PELa) was purified by affinity chromatography in a mannose-agarose column. The lectin agglutinated rabbit erythrocytes and the agglutinating effect was inhibited by previous incubation with the glycoprotein fetuin, along with N-acetyl-D-glucosamine, D-mannose and its derivatives. The lectin maintained complete activity in temperatures ranging from 40 to 60 C and pH values ranging from 9 to 10. As a glycoprotein, PELa has a carbohydrate content of 2.2%, and its activity requires divalent cations such as Ca2+ and Mn2+. Based on SDS-PAGE, PELa displays a profile similar to that of other Dalbergieae lectins with the main chain of molecular mass around 30 kDa and two subunits of 19 kDa and 10 kDa each. Two-dimensional (2D) electrophoresis revealed the presence of isoforms with different isoelectric points, and high-performance size exclusion chromatography (HPSEC) was performed to confirm the purity of the sample. The lectin was immobilized in CNBr-activated Sepharose 4B and successfully captured fetuin in solution, demonstrating that this lectin remains active and capable of binding carbohydrates. PELa showed effects different from those of its recombinant form in both pro and anti-inflammatory tests. (C) 2017 Elsevier B.V. All rights reserved.

引用

页码：323 / 330

页数：8

共 59 条

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