Yoo Jae Il, Kim Hwa Su, Choi Chi Won, Yoo Jung Sik, Yu Jae Yon, Lee Yeong Seon
Division of Antimicrobial Resistance, Korea National Institute of Health, Osong, Korea.
Proteome Research Team, Korea Basic Science Institute, Daejeon, Korea.
Osong Public Health Res Perspect. 2013 Dec;4(6):293-300. doi: 10.1016/j.phrp.2013.10.001. Epub 2013 Oct 12.
The proteomic analysis of voriconazole resistant Candida glabrata strain has not yet been investigated. In this study, differentially expressed proteins of intracellular and membrane fraction from voriconazole-susceptible, susceptible dose-dependent (S-DD), resistant C. glabrata strains were compared with each other and several proteins were identified.
The proteins of intracellular and membrane were isolated by disrupting cells with glass bead and centrifugation from voriconazole susceptible, S-DD, and resistant C. glabrata strains. The abundance of expressed proteins was compared using two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis and proteins showing continuous twofold or more increase or reduction of expression in resistant strains compared to susceptible and S-DD strain were analyzed by liquid chromatography/mass spectrometry-mass spectrometry method.
Of 34 intracellular proteins, 15 proteins showed expression increase or reduction (twofold or more). The identified proteins included regulation, energy production, carbohydrate transport, amino acid transport, and various metabolism related proteins. The increase of expression of heat shock protein 70 was found. Among membrane proteins, 12, 31 proteins showed expression increase or decrease in the order of susceptible, S-DD, and resistant strains. This expression included carbohydrate metabolism, amino acid synthesis, and response to stress-related proteins. In membrane fractions, the change of expression of 10 heat shock proteins was observed, and 9 heat shock protein 70 (Hsp70) showed the reduction of expression.
The expression of Hsp70 protein in membrane fraction is related to voriconazole resistant C. glabrata strains.
尚未对伏立康唑耐药光滑念珠菌菌株进行蛋白质组学分析。在本研究中，比较了伏立康唑敏感、敏感剂量依赖型（S-DD）、耐药光滑念珠菌菌株细胞内和膜部分的差异表达蛋白，并鉴定了几种蛋白。
通过玻璃珠破碎细胞并离心，从伏立康唑敏感、S-DD和耐药光滑念珠菌菌株中分离细胞内和膜蛋白。使用二维十二烷基硫酸钠聚丙烯酰胺凝胶电泳比较表达蛋白的丰度，对于与敏感和S-DD菌株相比在耐药菌株中表达持续增加或减少两倍或更多的蛋白，采用液相色谱/质谱-质谱法进行分析。
在34种细胞内蛋白中，有15种蛋白表达增加或减少（两倍或更多）。鉴定出的蛋白包括调节、能量产生、碳水化合物转运、氨基酸转运以及各种代谢相关蛋白。发现热休克蛋白70的表达增加。在膜蛋白中，分别有12种、31种蛋白在敏感、S-DD和耐药菌株中呈表达增加或减少。这种表达包括碳水化合物代谢、氨基酸合成以及应激相关蛋白。在膜部分，观察到10种热休克蛋白的表达变化，其中9种热休克蛋白70（Hsp70）表达降低。
膜部分中Hsp70蛋白的表达与伏立康唑耐药光滑念珠菌菌株有关。
