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Using mutagenesis and structural biology to map the binding site for the plasmodium falciparum merozoite Protein PfRh4 on the human immune adherence receptor
- Source :
- Park, H J, Guariento, M, Maciejewski, M, Hauhart, R, Tham, W-H, Cowman, A F, Schmidt, C Q, Mertens, H D T, Liszewski, M K, Hourcade, D E, Barlow, P N & Atkinson, J P 2014, ' Using mutagenesis and structural biology to map the binding site for the plasmodium falciparum merozoite Protein PfRh4 on the human immune adherence receptor ', Journal of Biological Chemistry, vol. 289, no. 1, pp. 450-463 . https://doi.org/10.1074/jbc.M113.520346
- Publication Year :
- 2014
-
Abstract
- To survive and replicate within the human host, malaria parasites must invade erythrocytes. Invasion can be mediated by the P. falciparum reticulocyte-binding homologue protein 4 (PfRh4) on the merozoite surface interacting with complement receptor type 1 (CR1, CD35) on the erythrocyte membrane. The PfRh4 attachment site lies within the three N-terminal complement control protein modules (CCPs 1-3) of CR1, which intriguingly also accommodate binding and regulatory sites for the key complement activation-specific proteolytic products, C3b and C4b. One of these regulatory activities is decay-accelerating activity. Although PfRh4 does not impact C3b/C4b binding, it does inhibit this convertase disassociating capability. Here, we have employed ELISA, co-immunoprecipitation, and surface plasmon resonance to demonstrate that CCP 1 contains all the critical residues for PfRh4 interaction. We fine mapped by homologous substitution mutagenesis the PfRh4-binding site on CCP 1 and visualized it with a solution structure of CCPs 1-3 derived by NMR and small angle x-ray scattering. We cross-validated these results by creating an artificial PfRh4-binding site through substitution of putative PfRh4-interacting residues from CCP 1 into their homologous positions within CCP 8; strikingly, this engineered binding site had an approximate to 30-fold higher affinity for PfRh4 than the native one in CCP 1. These experiments define a candidate site on CR1 by which P. falciparum merozoites gain access to human erythrocytes in a non-sialic acid-dependent pathway of merozoite invasion.
- Subjects :
- Plasmodium
Erythrocytes
MALARIA PARASITE INVASION
Protozoan Proteins
Complement System
Complement receptor
Biochemistry
Papua-New-Guinea
Protein structure
Glycophorin-C
X-Ray Diffraction
Complement Receptor Type 1
Ligand-Binding
Cell Surface Receptor
Complement C4b
Cell biology
Complement C3b
Protein Structure
Convertases
C3B/C4B receptor
Immunology
Plasmodium falciparum
Biology
complement receptor
parasitic diseases
Scattering, Small Angle
Humans
Binding site
Surface Plasmon Resonance (SPR)
Nuclear Magnetic Resonance, Biomolecular
Molecular Biology
Binding Sites
Merozoites
Immune adherence
Reticulocyte-binding Homologue Proteins
C3B binding
Membrane Proteins
Cell Biology
Surface Plasmon Resonance
Molecular biology
NMR
Complement system
Malaria
Erythrocyte invasion
HEK293 Cells
2 active-sites
Mutagenesis
Factor-H
Receptors, Complement 3b
biology.protein
Complement control protein
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Park, H J, Guariento, M, Maciejewski, M, Hauhart, R, Tham, W-H, Cowman, A F, Schmidt, C Q, Mertens, H D T, Liszewski, M K, Hourcade, D E, Barlow, P N & Atkinson, J P 2014, ' Using mutagenesis and structural biology to map the binding site for the plasmodium falciparum merozoite Protein PfRh4 on the human immune adherence receptor ', Journal of Biological Chemistry, vol. 289, no. 1, pp. 450-463 . https://doi.org/10.1074/jbc.M113.520346
- Accession number :
- edsair.doi.dedup.....8f51e4227fc08d67aa2943b933498d96