Lan You, Zhou Mao, Li Xin, Liu Xuan, Li Jun, Liu Wenen
Department of Clinical Laboratory, Xiangya Hospital Central South University, Changsha, Hunan, 410008, People's Republic of China.
Infect Drug Resist. 2022 Feb 2;15:311-320. doi: 10.2147/IDR.S342368. eCollection 2022.
To investigate the role of outer membrane vesicles (OMVs) and related proteins in iron acquisition of hypervirulent (HVKP) and classic (cKP).
The OMVs of HVKP and cKP under iron-deficient and iron-sufficient media were extracted and purified by ultracentrifugation. Transmission electron microscopy (TEM) was used to identify OMVs. The quantitative proteomics were performed based on mass spectrometry.
Four OMVs samples secreted by HVKP and cKP under iron-deficient and iron-sufficient environment were isolated and collected (HVKP OMVs under iron-deficient environment (A1), HVKP OMVs under iron-sufficient environment (A2), cKP OMVs under iron-deficient environment (B1), cKP OMVs under iron-sufficient environment (B2)). The amount of OMVs released by HVKP in iron-deficient medium was significantly larger than that in iron-sufficient medium ( < 0.05). HVKP secreted more OMVs than cKP in iron-deficient medium ( < 0.05). A total of 1074 kinds of proteins were identified in four samples. A comparison between the iron-deficient vs iron-sufficient environment showed that 61 proteins in HVKP OMVs were identified with a significant change in abundance under iron-deficient environment. Among them, 17 proteins were related to iron acquisition and transportation systems. While in cKP OMVs, 62 proteins significantly changed under iron-deficient environment in which 5 proteins were related to iron acquisition and transportation systems. Upon comparison of the HVKP vs cKP OMVs under iron deficiency, 81 proteins were detected with a significant change in which 8 proteins were related to iron acquisition and transportation systems.
Above all, the results of this study suggest a potential role for OMVs in iron acquisition of HVKP and provide evidence of potential connections between OMVs and strong iron-acquisition ability of HVKP during iron limitation.
探讨外膜囊泡（OMVs）及其相关蛋白在高毒力肺炎克雷伯菌（HVKP）和经典肺炎克雷伯菌（cKP）铁摄取中的作用。
采用超速离心法提取并纯化缺铁和富铁培养基条件下HVKP和cKP的OMVs。利用透射电子显微镜（TEM）鉴定OMVs。基于质谱进行定量蛋白质组学分析。
分离并收集了缺铁和富铁环境下HVKP和cKP分泌的4个OMVs样本（缺铁环境下的HVKP OMVs（A1）、富铁环境下的HVKP OMVs（A2）、缺铁环境下的cKP OMVs（B1）、富铁环境下的cKP OMVs（B2））。缺铁培养基中HVKP释放的OMVs量显著大于富铁培养基中的（<0.05）。缺铁培养基中HVKP分泌的OMVs比cKP多（<0.05）。4个样本中共鉴定出1074种蛋白质。缺铁与富铁环境比较显示，缺铁环境下HVKP OMVs中有61种蛋白质丰度有显著变化。其中，17种蛋白质与铁摄取和转运系统有关。而在cKP OMVs中，缺铁环境下有62种蛋白质显著变化，其中5种蛋白质与铁摄取和转运系统有关。缺铁条件下HVKP与cKP OMVs比较，检测到81种蛋白质有显著变化，其中8种蛋白质与铁摄取和转运系统有关。
综上所述，本研究结果提示OMVs在HVKP铁摄取中具有潜在作用，并为铁限制条件下OMVs与HVKP强大的铁摄取能力之间的潜在联系提供了证据。
