Department of Microbiology and Molecular Medicine, CMU, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland.
Department of Life Sciences, Imperial College London, South Kensington, London SW7 2AZ, UK.
Biochim Biophys Acta Proteins Proteom. 2017 Feb;1865(2):208-219. doi: 10.1016/j.bbapap.2016.11.010. Epub 2016 Nov 22.
The immune mapped protein 1 (IMP1) was first identified as a protective antigen in Eimeria maxima and described as vaccine candidate and invasion factor in Toxoplasma gondii. We show here that TgIMP1 localizes to the inner leaflet of plasma membrane (PM) via dual acylation. Mutations either in the N-terminal myristoylation or palmitoylation sites (G2 and C5) cause relocalization of TgIMP1 to the cytosol. The first 11 amino acids are sufficient for PM targeting and the presence of lysine (K7) is critical. Disruption of TgIMP1 gene by double homologous recombination revealed no invasion defect or any measurable alteration in the lytic cycle of tachyzoites. Following immunization with TgIMP1 DNA vaccine, mice challenged with either wild type or IMP1-ko parasites showed no significant difference in protection. The sequence analysis identified a structured C-terminal domain that is present in a broader family of IMP1-like proteins conserved across the members of Apicomplexa. We present the solution structure of this domain determined from NMR data and describe a new protein fold not seen before.
免疫映射蛋白 1(IMP1)最初在巨型艾美耳球虫中被鉴定为保护性抗原,并被描述为刚地弓形虫的疫苗候选物和入侵因子。我们在这里表明,TgIMP1 通过双重酰化定位在内质网的内层。在 N 端豆蔻酰化或棕榈酰化位点(G2 和 C5)的突变导致 TgIMP1 重新定位于细胞质。前 11 个氨基酸足以靶向质膜,赖氨酸(K7)的存在至关重要。通过双同源重组破坏 TgIMP1 基因,没有发现入侵缺陷或速殖子裂解周期有任何可测量的改变。用 TgIMP1 DNA 疫苗免疫后,用野生型或 IMP1-ko 寄生虫攻击的小鼠在保护方面没有明显差异。序列分析鉴定出一个结构域,该结构域存在于广泛的 IMP1 样蛋白家族中,在顶复门的成员中保守。我们从 NMR 数据确定了该结构域的溶液结构,并描述了一种以前未见的新蛋白折叠。