Heterologous expression and molecular binding properties of AofleA, a fucose-specific lectin from nematophagous fungus Arthrobotrys oligospora

Int J Biol Macromol. 2020 Apr 15:149:1213-1221. doi: 10.1016/j.ijbiomac.2020.02.039. Epub 2020 Feb 6.

Abstract

Lectins are the primary recognition macromolecules for various types of fucosylation, a common eukaryotic post-translational modification. In this study, we report the heterologous expression and molecular binding properties of a fucose-specific lectin, AofleA, isolated from Arthrobotrys oligospora. This is the first reported fucose-specific lectin found in nematophagous fungi. The recombinant AofleA (r-AofleA) was expressed in Escherichia coli with high efficiency, yielding at least 500 mg of soluble and functional r-AofleA per liter of broth. Using hemagglutination inhibition assay and glycan microarray analysis, r-AofleA was found to be broadly specific for fucosylated glycans or oligosaccharides including Fucα(1-2), Fucα(1-3), Fucα(1-4) and Fucα(1-6) linkages, similar to Aleuria aurantia lectin (AAL). Frontal affinity chromatography showed that r-AofleA has high affinity towards PA-L-fucose with an average Kd value of 15 nM. These findings provide a basis for improved understanding of the structure and functions of AofleA during recognition and capture of prey nematodes by nematophagous fungus A. oligospora.

Keywords: Arthrobotrys oligospora; Fucose-specific lectins; Glycan microarray analysis; Nematophagous fungi.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Ascomycota / metabolism*
  • Biological Assay
  • Erythrocytes / metabolism
  • Fungal Proteins / chemistry
  • Fungal Proteins / metabolism*
  • Hemagglutination
  • Kinetics
  • Lectins / chemistry
  • Lectins / metabolism*
  • Microarray Analysis
  • Models, Molecular
  • Plasmids / genetics
  • Polysaccharides / metabolism
  • Rabbits
  • Recombinant Proteins / isolation & purification

Substances

  • Fungal Proteins
  • Lectins
  • Polysaccharides
  • Recombinant Proteins
  • fucose-binding lectin