The knob protein KAHRP assembles into a ring-shaped structure that underpins virulence complex assembly

Oliver Looker, Adam J. Blanch, Boyin Liu, Juan Nunez-Iglesias, Paul McMillan, Leann Tilley, Matthew W A Dixon

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

Plasmodium falciparum mediates adhesion of infected red blood cells (RBCs) to blood vessel walls by assembling a multi-protein complex at the RBC surface. This virulence-mediating structure, called the knob, acts as a scaffold for the presentation of the major virulence antigen, P. falciparum Erythrocyte Membrane Protein-1 (PfEMP1). In this work we developed correlative STochastic Optical Reconstruction Microscopy–Scanning Electron Microscopy (STORM-SEM) to spatially and temporally map the delivery of the knob-associated histidine-rich protein (KAHRP) and PfEMP1 to the RBC membrane skeleton. We show that KAHRP is delivered as individual modules that assemble in situ, giving a ring-shaped fluorescence profile around a dimpled disk that can be visualized by SEM. Electron tomography of negatively-stained membranes reveals a previously observed spiral scaffold underpinning the assembled knobs. Truncation of the C-terminal region of KAHRP leads to loss of the ring structures, disruption of the raised disks and aberrant formation of the spiral scaffold, pointing to a critical role for KAHRP in assembling the physical knob structure. We show that host cell actin remodeling plays an important role in assembly of the virulence complex, with cytochalasin D blocking knob assembly. Additionally, PfEMP1 appears to be delivered to the RBC membrane, then inserted laterally into knob structures.
Original languageEnglish
Article numbere1007761
Number of pages26
JournalPLoS Pathogens
Volume15
Issue number5
DOIs
Publication statusPublished - 9 May 2019
Externally publishedYes

Keywords

  • scanning electron microscopy
  • parasitic diseases
  • spectrins
  • fluorescence imaging
  • actins
  • fluorescence
  • microbeads
  • protein structure

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