PMID: 9664038Jul 17, 1998Paper

The amphipathic alpha helices of the toxoplasma protein GRA2 mediate post-secretory membrane association

Journal of Cell Science
C MercierL D Sibley

Abstract

The Toxoplasma gondii protein GRA2 is secreted into the parasite-containing vacuole where it is rapidly and specifically targeted to a network of membranous tubules that connect with the vacuolar membrane. To examine the molecular basis of this association, we expressed an HA9 epitope-tagged form of GRA2 by stable transformation of Toxoplasma. GRA2-HA9 was correctly packaged inside the dense granules, secreted into the PV and targeted to the network, as shown by immunoelectron microscopy, immunofluorescence and cell fractionation. Expression of deletion mutants of GRA2-HA9 lacking either of two amphipathic alpha helices resulted in the production and secretion of soluble proteins which were unable to stably associate with the network. A mutant in which the amino acids of the first alpha helix were rearranged to a non-amphipathic pattern localized correctly to the network but failed to remained stably associated with the membrane. Collectively, these results demonstrate that targeting and membrane association occur by separate mechanisms and that the combination of both alpha helices is essential for stable localization of GRA2 to the network.

Citations

Jun 28, 2005·International Journal for Parasitology·Corinne MercierMarie-France-Cesbron Delauw
Mar 20, 2001·Molecular and Biochemical Parasitology·B StriepenD S Roos
Mar 18, 1999·Molecular and Biochemical Parasitology·A R BlancoR E Sinden
Sep 6, 2000·Current Opinion in Microbiology·A J Kaasch, K A Joiner
Apr 10, 1999·Molecular Biology of the Cell·L LecordierM F Cesbron-Delauw
Jul 23, 2002·Molecular Biology of the Cell·Corinne MercierMarie-France Cesbron-Delauw
Sep 7, 2000·Microbiology and Molecular Biology Reviews : MMBR·M W Black, J C Boothroyd
Sep 21, 2001·Biochemical and Biophysical Research Communications·H J AhnH W Nam
Jul 25, 2009·International Journal for Parasitology·John C Boothroyd
Feb 28, 2015·International Journal for Parasitology·Louis-Philippe LerouxFlorence S Dzierszinski
Apr 25, 2017·Cellular Microbiology·Kaice A LaFaversGustavo Arrizabalaga
May 20, 2006·Anais Da Academia Brasileira De Ciências·Wanderley de Souza

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