5KRP

Frutapin, a lectin from Artocarpus incisa: Cloning, Expressing and Structural Analysis.

Summary for 5KRP

• Entry DOI: 10.2210/pdb5krp/pdb
• Descriptor: Frutapin, GLYCEROL (3 entities in total)
• Functional Keywords: protein, lectin, apofrutapin, artocarpus altillis, carbohydrate binding protein
• Biological source: Artocarpus altilis
• Total number of polymer chains: 4
• Total formula weight: 66366.31

Authors

de Sousa, F.D.,Coker, A.R.,Guan, Y.,Guo, J.,de Oliveira Monteiro-Moreira, A.C.,de Azevedo Moreira, R. (deposition date: 2016-07-07, release date: 2017-07-19, Last modification date: 2023-10-04)

Primary citation

de Sousa, F.D.,da Silva, B.B.,Furtado, G.P.,Carneiro, I.S.,Lobo, M.D.P.,Guan, Y.,Guo, J.,Coker, A.R.,Lourenzoni, M.R.,Guedes, M.I.F.,Owen, J.S.,Abraham, D.J.,Monteiro-Moreira, A.C.O.,Moreira, R.A.
Frutapin, a lectin fromArtocarpus incisa(breadfruit): cloning, expression and molecular insights.
Biosci. Rep., 37:-, 2017

PubMed Abstract: (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 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 (P222), 1.70 (P321) and 1.60 (P321) 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.

PubMed: 28684550
DOI: 10.1042/BSR20170969

Experimental method

X-RAY DIFFRACTION (1.58 Å)