Putative membrane proteins identified in T. vaginalis hydrogenosomes (continued).

Proteins were manually annotated based on searches in TrichDB, Uniprot, and PFAM A+B (Table S2). Protein structure was predicted using TMHMM and MEMSAT3; subcellular location was predicted using TargetP and PsortII. TM No., number of predicted transmembrane α-helixes. M, predicted location in mitochondria. S - predicted proteins of secretory pathway; O, predicted location in other compartments; Mit%, probability percentage of mitochondrial location; *, mitochondrial location was not predicted; Exp. Local., experimental location; H, localization of HA-tagged proteins was confirmed in T. vaginalis hydrogenosomes by immunofluorescence microscopy; NV, transformed T. vaginalis strain was not viable. Signal: Δ indicates N-terminal targeting sequence identified by Hunter; β indicates presence of beta signal of beta-barrel proteins for insertion into the outer membrane of mitochondria; Ct, C-tail anchor detected [Fig. S2].

本研究基于TrichDB、通用蛋白质知识库（UniProt）及PFAM A+B数据库的检索结果，对蛋白质进行人工注释（详见补充表S2）。蛋白质结构预测采用TMHMM与MEMSAT3工具完成，亚细胞定位预测则通过TargetP和PsortII工具实现。相关参数与符号说明如下：TM编号（TM No.）：预测获得的跨膜α螺旋数量；M：预测定位于线粒体；S：预测属于分泌通路的蛋白质；O：预测定位于其他细胞区室；Mit%：线粒体定位的概率百分比；*：未预测到线粒体定位；Exp. Local.：实验验证的定位信息；H：通过免疫荧光显微镜术证实，HA标签（HA-tag）融合蛋白在阴道毛滴虫（T. vaginalis）氢化酶体中的定位；NV：转化后的阴道毛滴虫菌株无法存活；信号说明：Δ代表Hunter鉴定得到的N端靶向序列；β代表β桶状蛋白所携带的、可介导其插入线粒体外膜的β信号序列；Ct：检测到C端锚定序列[详见补充图S2]。