title: Unusual N-glycan structures required for trafficking Toxoplasma gondii GAP50 to the inner membrane complex regulate host cell entry through parasite motility.
abstract: Toxoplasma gondii motility, which is essential for host cell entry, migration through host tissues, and invasion, is a unique form of actin-dependent gliding. It is powered by a motor complex mainly composed of myosin heavy chain A, myosin light chain 1, gliding associated proteins GAP45, and GAP50, the only integral membrane anchor so far described. In the present study, we have combined glycomic and proteomic approaches to demonstrate that all three potential N-glycosylated sites of GAP50 are occupied by unusual N-glycan structures that are rarely found on mature mammalian glycoproteins. Using site-directed mutagenesis, we show that N-glycosylation is a prerequisite for GAP50 transport from the endoplasmic reticulum to the Golgi apparatus and for its subsequent delivery into the inner complex membrane. Assembly of key partners into the gliding complex, and parasite motility are severely impaired in the unglycosylated GAP50 mutants. Furthermore, comparative affinity purification using N-glycosylated and unglycosylated GAP50 as bait identified three novel hypothetical proteins including the recently described gliding associated protein GAP40, and we demonstrate that N-glycans are required for efficient binding to gliding partners. Collectively, these results provide the first detailed analyses of T. gondii N-glycosylation functions that are vital for parasite motility and host cell entry.
authors: Center for Infection and Immunity of Lille, CNRS UMR 8204, INSERM U 1019, Institut Pasteur de Lille, Université Lille Nord de France, 59000 Lille, France.
authors: Sylvain
authors: S
authors: Fauquenoy
authors: Agnès
authors: A
authors: Hovasse
authors: Pierre-Julien
authors: PJ
authors: Sloves
authors: Willy
authors: W
authors: Morelle
authors: Tchilabalo
authors: T
authors: Dilezitoko Alayi
authors: Tchilabalo
authors: T
authors: Dilezitoko Ayali
authors: Christian
authors: C
authors: Slomianny
authors: Elisabeth
authors: E
authors: Werkmeister
authors: Christine
authors: C
authors: Schaeffer
authors: Alain
authors: A
authors: Van Dorsselaer
authors: Stanislas
authors: S
authors: Tomavo
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