title: The 10 kDa domain of human erythrocyte protein 4.1 binds the Plasmodium falciparum EBA-181 protein.
abstract: BACKGROUND
  Erythrocyte invasion by Plasmodium falciparum parasites represents a key mechanism during malaria pathogenesis. Erythrocyte binding antigen-181 (EBA-181) is an important invasion protein, which mediates a unique host cell entry pathway. A novel interaction between EBA-181 and human erythrocyte membrane protein 4.1 (4.1R) was recently demonstrated using phage display technology. In the current study, recombinant proteins were utilized to define and characterize the precise molecular interaction between the two proteins.
  
  
  METHODS
  4.1R structural domains (30, 16, 10 and 22 kDa domain) and the 4.1R binding region in EBA-181 were synthesized in specific Escherichia coli strains as recombinant proteins and purified using magnetic bead technology. Recombinant proteins were subsequently used in blot-overlay and histidine pull-down assays to determine the binding domain in 4.1R.
  
  
  RESULTS
  Blot overlay and histidine pull-down experiments revealed specific interaction between the 10 kDa domain of 4.1R and EBA-181. Binding was concentration dependent as well as saturable and was abolished by heat denaturation of 4.1R.
  
  
  CONCLUSIONS
  The interaction of EBA-181 with the highly conserved 10 kDa domain of 4.1R provides new insight into the molecular mechanisms utilized by P. falciparum during erythrocyte entry. The results highlight the potential multifunctional role of malaria invasion proteins, which may contribute to the success of the pathogenic stage of the parasite's life cycle.
authors.affiliation: Department of Molecular Medicine and Haematology, National Health Laboratory Service, School of Pathology, University of the Witwatersrand, Parktown, Johannesburg, 2193, South Africa. roberto.lanzillotti@gmail.com
authors.forename: Roberto
authors.initials: R
authors.lastname: Lanzillotti
authors.forename: Theresa L
authors.initials: TL
authors.lastname: Coetzer
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism / Q000469:parasitology; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000378:metabolism / Q000472:pathogenicity*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D000953:Antigens, Protozoan; D003598:Cytoskeletal Proteins; C474029:EBA-181 protein, Plasmodium falciparum; D008565:Membrane Proteins; D011994:Recombinant Proteins; C028841:erythrocyte membrane band 4.1 protein