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Frutapin, a lectin from Artocarpus incisa (breadfruit): cloning, expression and molecular insights.

Abstract：

:Artocarpus incisa (breadfruit) seeds contain three different lectins (Frutalin, Frutapin (FTP) and Frutackin) with distinct carbohydrate specificities. The most abundant lectin is Frutalin, an α-D-galactose-specific carbohydrate-binding glycoprotein with antitumour properties and potential for tumour biomarker discovery as already reported. FTP is the second most abundant, but proved difficult to purify with very low yields and contamination with Frutalin frustrating its characterization. Here, we report for the first time high-level production and isolation of biologically active recombinant FTP in Escherichia coli BL21, optimizing conditions with the best set yielding >40 mg/l culture of soluble active FTP. The minimal concentration for agglutination of red blood cells was 62.5 µg/ml of FTP, a process effectively inhibited by mannose. Apo-FTP, FTP-mannose and FTP-glucose crystals were obtained, and they diffracted X-rays to a resolution of 1.58 (P212121), 1.70 (P3121) and 1.60 (P3121) Å respectively. The best solution showed four monomers per asymmetric unit. Molecular dynamics (MD) simulation suggested that FTP displays higher affinity for mannose than glucose. Cell studies revealed that FTP was non-cytotoxic to cultured mouse fibroblast 3T3 cells below 0.5 mg/ml and was also capable of stimulating cell migration at 50 µg/ml. In conclusion, our optimized expression system allowed high amounts of correctly folded soluble FTP to be isolated. This recombinant bioactive lectin will now be tested in future studies for therapeutic potential; for example in wound healing and tissue regeneration.

journal_name

Biosci Rep

journal_title

Bioscience reports

authors

de Sousa FD,da Silva BB,Furtado GP,Carneiro IS,Lobo MDP,Guan Y,Guo J,Coker AR,Lourenzoni MR,Guedes MIF,Owen JS,Abraham DJ,Monteiro-Moreira ACO,Moreira RA

doi

10.1042/BSR20170969

subject

Has Abstract

pub_date

2017-07-21 00:00:00

issue

4

eissn

0144-8463

issn

1573-4935

pii

BSR20170969

journal_volume

37

pub_type

杂志文章

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