Solution structure, glycan specificity and of phenol oxidase inhibitory activity of Anopheles C-type lectins CTL4 and CTLMA2

Sci Rep. 2019 Oct 23;9(1):15191. doi: 10.1038/s41598-019-51353-z.

Abstract

Malaria, the world's most devastating parasitic disease, is transmitted between humans by mosquitoes of the Anopheles genus. An. gambiae is the principal malaria vector in Sub-Saharan Africa. The C-type lectins CTL4 and CTLMA2 cooperatively influence Plasmodium infection in the malaria vector Anopheles. Here we report the purification and biochemical characterization of CTL4 and CTLMA2 from An. gambiae and An. albimanus. CTL4 and CTLMA2 are known to form a disulfide-bridged heterodimer via an N-terminal tri-cysteine CXCXC motif. We demonstrate in vitro that CTL4 and CTLMA2 intermolecular disulfide formation is promiscuous within this motif. Furthermore, CTL4 and CTLMA2 form higher oligomeric states at physiological pH. Both lectins bind specific sugars, including glycosaminoglycan motifs with β1-3/β1-4 linkages between glucose, galactose and their respective hexosamines. Small-angle x-ray scattering data supports a compact heterodimer between the CTL domains. Recombinant CTL4/CTLMA2 is found to function in vivo, reversing the enhancement of phenol oxidase activity in dsCTL4-treated mosquitoes. We propose these molecular features underline a common function for CTL4/CTLMA2 in mosquitoes, with species and strain-specific variation in degrees of activity in response to Plasmodium infection.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Anopheles / metabolism*
  • Calcium / metabolism
  • Conserved Sequence
  • Enzyme Inhibitors / pharmacology*
  • Escherichia coli / metabolism
  • Insect Proteins / chemistry*
  • Insect Proteins / metabolism*
  • Lectins, C-Type / chemistry*
  • Lectins, C-Type / metabolism*
  • Monophenol Monooxygenase / antagonists & inhibitors*
  • Monophenol Monooxygenase / metabolism
  • Polysaccharides / metabolism*
  • Recombinant Proteins / metabolism
  • Solutions

Substances

  • Enzyme Inhibitors
  • Insect Proteins
  • Lectins, C-Type
  • Polysaccharides
  • Recombinant Proteins
  • Solutions
  • Monophenol Monooxygenase
  • Calcium