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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

Park, HJ, Guariento, M, Maciejewski, M, Hauhart, R, Tham, W, Cowman, AF, Schmidt, CQ, Mertens, HDT, Kathryn, LM, Hourcade, DE, Barlow, PN and Atkinson, JP (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, 289 (1). pp. 450-463. ISSN 0021-92581083-351X


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 crossvalidated 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 ~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. 2014 by The American Society for Biochemistry and Molecular Biology, Inc

Item Type: Article
Additional Information: NIBR author: Maciejewski, M institute: NIBR- address only contributor address: (Park, Hauhart, Kathryn Liszewski, Hourcade, Atkinson) Washington University, Division of Rheumatology, Department of Internal Medicine, 660 South Euclid, Campus Box 8045, St. Louis, MO 63110, United States (Guariento, Maciejewski, Schmidt, Barlow) School of Chemistry and Biological Sciences, University of Edinburgh, Edinburgh EH93JJ, United Kingdom (Tham, Cowman) Department of Medical Biology, Division of Infection and Immunity, University of Melbourne, Parkville, VIC 3052, Australia (Tham, Cowman) Walter and Eliza Hall Institute, Parkville, VIC 3052, Australia (Mertens) Australian National Synchrotron, Melbourne 3168, Australia (Park) Emkey Arthritis Clinic, 1200 Broadcasting Rd., Wyomissing, PA 19610, United States (Guariento) Friedrich Miescher Laboratory of the Max Planck Society, Spemannstr. 39, 72076 Tuebingen, Germany (Maciejewski) Novartis Institute for Biomedical Research, 250 Massachusetts Ave., Cambridge, MA 02139, United States (Schmidt) Institute of Pharmacology of Natural Products and Clinical Pharmacology, Ulm University, Helmholtzstr. 20, D-89081 Ulm, Germany
Date Deposited: 13 Oct 2015 13:12
Last Modified: 04 Jul 2016 23:45