The fucomic potential of mosquitoes: Fucosylated N-glycan epitopes and their cognate fucosyltransferases

Simone Kurz, Jonas G. King, Rhoel R. Dinglasan, Katharina Paschinger, Iain B.H. Wilson

Research output: Contribution to journalArticle

Abstract

Fucoconjugates are key mediators of protein-glycan interactions in prokaryotes and eukaryotes. As examples, N-glycans modified with the non-mammalian core α1,3-linked fucose have been detected in various organisms ranging from plants to insects and are immunogenic in mammals. The rabbit polyclonal antibody raised against plant horseradish peroxidase (anti-HRP) is able to recognize the α1,3-linked fucose epitope and is also known to specifically stain neural tissues in the fruit fly Drosophila melanogaster. In this study, we have detected and localized the anti-HRP cross-reactivity in another insect species, the malaria mosquito vector Anopheles gambiae. We were able to identify and structurally elucidate fucosylated N-glycans including core mono- and difucosylated structures (responsible for anti-HRP cross reactivity) as well as a Lewis-type antennal modification on mosquito anionic N-glycans by applying enzymatic and chemical treatments. The three mosquito fucosyltransferase open reading frames (FucT6, FucTA and FucTC) required for the in vivo biosynthesis of the fucosylated N-glycan epitopes were identified in the Anopheles gambiae genome, cloned and recombinantly expressed in Pichia pastoris. Using a robust MALDI-TOF MS approach, we characterised the activity of the three recombinant fucosyltransferases in vitro and demonstrate that they share similar enzymatic properties as compared to their homologues from D. melanogaster and Apis mellifera. Thus, not only do we confirm the neural reactivity of anti-HRP in a mosquito species, but also demonstrate enzymatic activity for all its α1,3- and α1,6-fucosyltransferase homologues, whose specificity matches the results of glycomic analyses.

Original languageEnglish (US)
Pages (from-to)52-63
Number of pages12
JournalInsect Biochemistry and Molecular Biology
Volume68
DOIs
StatePublished - Jan 1 2016

Keywords

  • Anopheles gambiae
  • Anti-HRP
  • Fucosyltransferases
  • Immunofluorescence
  • Mass spectrometry
  • N-glycans

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Insect Science

Fingerprint Dive into the research topics of 'The fucomic potential of mosquitoes: Fucosylated N-glycan epitopes and their cognate fucosyltransferases'. Together they form a unique fingerprint.

  • Cite this