title: Structure-function analysis of the Anopheles gambiae LRIM1/APL1C complex and its interaction with complement C3-like protein TEP1.
abstract: Malaria threatens half the world's population and exacts a devastating human toll. The principal malaria vector in Africa, the mosquito Anopheles gambiae, encodes 24 members of a recently identified family of leucine-rich repeat proteins named LRIMs. Two members of this family, LRIM1 and APL1C, are crucial components of the mosquito complement-like pathway that is important for immune defense against Plasmodium parasites. LRIM1 and APL1C circulate in the hemolymph exclusively as a disulfide-bonded complex that specifically interacts with the mature form of the complement C3-like protein, TEP1. We have investigated the specificity of LRIM1/APL1C complex formation and which regions of these proteins are required for interactions with TEP1. To address these questions, we have generated a set of LRIM1 and APL1C alleles altering key conserved structural elements and assayed them in cell culture for complex formation and interaction with TEP1. Our data indicate that heterocomplex formation is an intrinsic ability of LRIM1 and APL1C and identify key homologous cysteine residues forming the intermolecular disulfide bond. We also demonstrate that the coiled-coil domain is the binding site for TEP1 but also contributes to the specificity of LRIM1/APL1C complex formation. In addition, we show that the LRIM1/APL1C complex interacts with the mature forms of three other TEP proteins, one of which, TEP3, we have characterized as a Plasmodium antagonist. We conclude that LRIM1 and APL1C contain three distinct modules: a C-terminal coiled-coil domain that can carry different TEP protein cargoes, potentially with distinct functions, a central cysteine-rich region that controls complex formation and an N-terminal leucine-rich repeat with a putative role in pathogen recognition.
authors.affiliation: Division of Cell and Molecular Biology, Department of Life Sciences, Imperial College, London, United Kingdom.
authors.forename: Michael
authors.initials: M
authors.lastname: Povelones
authors.forename: Leanna M
authors.initials: LM
authors.lastname: Upton
authors.forename: Katarzyna A
authors.initials: KA
authors.lastname: Sala
authors.forename: George K
authors.initials: GK
authors.lastname: Christophides
mesh_terms: D000818:Animals; D000852:Anopheles / Q000235:genetics / Q000378:metabolism* / Q000469:parasitology; D006458:Hemolymph / Q000378:metabolism / Q000469:parasitology; D006801:Humans; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D008288:Malaria / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D010962:Plasmodium berghei / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship
chemical_list: D019476:Insect Proteins; C540280:LRIM1 protein, Anopheles gambiae; D046912:Multiprotein Complexes; C544265:TEP1 protein, Anopheles gambiae