Isoform Characterisation, Heterologous Expression and Functional Analysis of Two Lectins from Vatairea macrocarpa

Author(s): João Garcia Alves Filho, Antonia Sâmia Fernandes do Nascimento, Ana Cláudia Silva Gondim, Ronniery Hilario Pereira, Rodrigo Maranguape Silva da Cunha, Celso Shiniti Nagano, Edson Holanda Teixeira, Kyria Santiago Nascimento, Benildo Sousa Cavada.

Journal Name: Protein & Peptide Letters

Volume 20 , Issue 11 , 2013

Become EABM
Become Reviewer

Abstract:

VML is a lectin from Vatairea macrocarpa seeds that has various biological activities. Here, we describe three new lectin isoforms from V. macrocarpa identified through genomic DNA analysis. One of these isoforms has high similarity to VML, while another that has noteworthy differences. We have denoted the new isoforms as VML-2, VML-3 and VML-4. Recombinant VML (rVML) and VML-2 (rVML-2) were expressed in Escherichia coli and were anticipated to have similar biological activity compared to native VML. Recombinant lectins were produced using a synthetic gene strategy to improve the expression levels. We obtained two active recombinant lectin isoforms from V. macrocarpa, and there was no significant difference between their biological activities. The conservation between carbohydrate-binding sites of recombinant and native proteins was demonstrated by specific inhibition of hemagglutin activity by D-galactose and lactose. However, no inhibition was observed in the presence of glucose and mannose. Our data show that the recombinant lectins VML and VML-2 are active and capable of recognising D-galactose and lactose. Moreover, the absence of glycosylation does not interfere with their biological activity.

Keywords: Active protein, heterologous expression, isoform, lectin, legume, recombinant.

Rights & PermissionsPrintExport Cite as

Article Details

VOLUME: 20
ISSUE: 11
Year: 2013
Page: [1204 - 1210]
Pages: 7
DOI: 10.2174/09298665113209990049
Price: $65

Article Metrics

PDF: 9

Special-new-year-discount