Wang Wenbin, Liu Jianxin, Guo Shanshan, Liu Lei, Yuan Qianyun, Guo Lei, Pan Saikun
Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang, China.
Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Ocean University, Lianyungang, China.
Front Microbiol. 2021 Jan 28;11:625315. doi: 10.3389/fmicb.2020.625315. eCollection 2020.
The discovery of outer membrane proteins (OMPs) with desirable specificity and surface availability is a fundamental challenge to develop accurate immunodiagnostic assay and multivalent vaccine of pathogenic species in food and aquaculture. Herein 101 OMPs were systemically screened from 4,831 non-redundant proteins of by bioinformatical predication of signaling peptides, transmembrane (TM) α-helix, and subcellular location. The sequence homology analysis with 32 species of spp. and all the non- strains revealed that 15 OMPs were conserved in at least 23 species, including BamA (VP2310), GspD (VP0133), Tolc (VP0425), OmpK (VP2362), OmpW (VPA0096), LptD (VP0339), Pal (VP1061), flagellar L-ring protein (VP0782), flagellar protein MotY (VP2111), hypothetical protein (VP1713), fimbrial assembly protein (VP2746), VacJ lipoprotein (VP2214), agglutination protein (VP1634), and lipoprotein (VP1267), Chitobiase (VP0755); high adhesion probability of flgH, LptD, OmpK, and OmpW indicated they were potential multivalent vaccine candidates. OMPs were found to share high homology with at least one or two species, 19 OMPs including OmpA like protein (VPA073), CsuD (VPA1504), and MtrC (VP1220) were found relatively specific to . The surface proteomic study by enzymatical shaving the cells showed the capsular polysaccharides most likely limited the protease action, while the glycosidases improved the availability of OMPs to trypsin. The OmpA (VPA1186, VPA0248, VP0764), Omp (VPA0166), OmpU (VP2467), BamA (VP2310), TolC (VP0425), GspD (VP0133), OmpK (VP2362), lpp (VPA1469), Pal (VP1061), agglutination protein (VP1634), and putative iron (III) compound receptor (VPA1435) have better availability on the cell surface.
发现具有理想特异性和表面可及性的外膜蛋白（OMPs）是开发针对食品和水产养殖中致病物种的准确免疫诊断检测方法和多价疫苗的一项基本挑战。在此，通过对信号肽、跨膜（TM）α螺旋和亚细胞定位进行生物信息学预测，从4831个非冗余蛋白中系统筛选出了101个OMPs。与32种嗜水气单胞菌属物种和所有非嗜水气单胞菌菌株的序列同源性分析表明，15个OMPs在至少23种物种中保守，包括BamA（VP2310）、GspD（VP0133）、TolC（VP0425）、OmpK（VP2362）、OmpW（VPA0096）、LptD（VP0339）、Pal（VP1061）、鞭毛L环蛋白（VP0782）、鞭毛蛋白MotY（VP2111）、假定蛋白（VP1713）、菌毛组装蛋白（VP2746）、VacJ脂蛋白（VP22,14）、凝集蛋白（VP1634）、脂蛋白（VP1267）、壳二糖酶（VP0755）；flgH、LptD、OmpK和OmpW的高黏附概率表明它们是潜在的多价疫苗候选物。发现OMPs与至少一种或两种嗜水气单胞菌属物种具有高度同源性，19个OMPs包括OmpA样蛋白（VPA073）、CsuD（VPA1504）和MtrC（VP1220）被发现对嗜水气单胞菌属相对特异。通过酶解细胞进行的表面蛋白质组学研究表明，荚膜多糖很可能限制了蛋白酶的作用，而糖苷酶提高了OMPs对胰蛋白酶的可及性。OmpA（VPA1186、VPA0248、VP0764）、Omp（VPA0166）、OmpU（VP2467）、BamA（VP2310）、TolC（VP0425）、GspD（VP0133）、OmpK（VP2362）、lpp（VPA1469）、Pal（VP1061）、凝集蛋白（VP1634）和假定的铁（III）化合物受体（VPA1435）在细胞表面具有更好的可及性。
