Toxoplasma gondii: Biochemical and biophysical characterization of recombinant soluble dense granule proteins GRA2 and GRA6

被引：12

作者：

Bittame, Amina ^{[1
,2
]}

Effantin, Gregory ^{[3
,4
,5
,6
]}

Petre, Graciane ^{[1
,2
]}

Ruffiot, Pauline ^{[1
,2
]}

Travier, Laetitia ^{[1
,2
]}

Schoehn, Guy ^{[3
,4
,5
,6
]}

Weissenhorn, Winfried ^{[3
,4
,5
,6
]}

Cesbron-Delauw, Marie-France ^{[1
,2
]}

Gagnon, Jean ^{[1
,2
]}

Mercier, Corinne ^{[1
,2
]}

机构：

[1] CNRS, UMR 5163, F-38042 Grenoble, France

[2] Univ Grenoble Alpes, F-38042 Grenoble, France

[3] Univ Grenoble Alpes, Inst Biol Struct, F-38044 Grenoble, France

[4] CNRS, IBS, F-38044 Grenoble, France

[5] CEA, IBS, F-38044 Grenoble, France

[6] UMI 3265 UFJ EMBL CNRS, Unit Virus Host Cell Interact, F-38027 Grenoble, France

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2015年 / 459卷 / 01期

关键词：

Amphipathic alpha-helices; Dense granule proteins (GRA); Toxoplasma gondii; Circular dichroism; Dynamic light scattering; Transmission electron microscopy; AMINO-ACID-SEQUENCE; PARASITOPHOROUS VACUOLE; HOST-CELL; INTRAVACUOLAR NETWORK; NANOTUBULAR NETWORK; MEMBRANE; PURIFICATION; SECRETION; DOMAIN; HELIX;

D O I：

10.1016/j.bbrc.2015.02.078

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The most prominent structural feature of the parasitophorous vacuole (PV) in which the intracellular parasite Toxoplasma gondii proliferates is a membranous nanotubular network (MNN), which interconnects the parasites and the PV membrane. The MNN function remains unclear. The GRA2 and GRA6 proteins secreted from the parasite dense granules into the PV have been implicated in the MNN biogenesis. Amphipathic alpha-helices (AAHs) predicted in GRA2 and an alpha-helical hydrophobic domain predicted in GRA6 have been proposed to be responsible for their membrane association, thereby potentially molding the MMN in its structure. Here we report an analysis of the recombinant proteins (expressed in detergent-free conditions) by circular dichroism, which showed that full length GRA2 displays an alpha-helical secondary structure while recombinant GRA6 and GRA2 truncated of its AAHs are mainly random coiled. Dynamic light scattering and transmission electron microscopy showed that recombinant GRA6 and truncated GRA2 constitute a homogenous population of small particles (6-8 nm in diameter) while recombinant GRA2 corresponds to 2 populations of particles (similar to 8-15 nm and up to 40 nm in diameter, respectively). The unusual properties of GRA2 due to its AAHs are discussed. (C) 2015 Elsevier Inc. All rights reserved.

引用

页码：107 / 112

页数：6

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