Analyzing the Hydrophobic Proteome of the Antarctic Archaeon Methanococcoides burtonii Using Differential Solubility Fractionation
Analyzing the Hydrophobic Proteome of the Antarctic Archaeon Methanococcoides burtonii Using Differential Solubility Fractionation
复制标题
DOI：
10.1021/pr9007865
复制
发表时间：
2010-02-01
影响因子：
4.4
通讯作者：
Cavicchioli, Ricardo
中科院分区：
文献类型：
--
作者：
Burg, Dominic W.;Lauro, Federico M.;Cavicchioli, Ricardo
蛋白质组学研究已被证明是有用的研究南极古菌Methanococcoides burtonii;然而，很少有人知道的疏水和膜蛋白，尽管他们的生物学重要性的知识。在这项研究中，开发了新的方法来分析和最大化的疏水蛋白质组的覆盖范围。分析的核心是使用正辛基-β-D-吡喃葡萄糖苷的微分溶解度分级（DSF）程序。该研究实现了已知表达蛋白总数的显著增加（330）。从鉴定的612个中，预测185个含有跨膜结构域或与膜相关，190个是疏水性的。DSF程序将识别膜蛋白的效率提高了169%，并且是经济的，与没有DSF的程序相比，需要更少的运行（12%的机器时间）来分析蛋白质组。肽光谱计数的分析使得能够评估生长温度特异性蛋白质。这种半定量分析是特别有用的鉴定低丰度的蛋白质不能使用标记策略进行定量。新鉴定的蛋白质的蛋白质组学分析揭示了许多以前与细胞适应无关的细胞过程，这种基于DSF的方法可能有利于广泛的生物系统的疏水蛋白质组学分析。
Proteomic studies have proven useful for studying the Antarctic archaeon Methanococcoides burtonii; however, little has been learned about the hydrophobic and membrane proteins, despite knowledge of their biological importance. In this study, new methods were developed to analyze and maximize the coverage of the hydrophobic proteome. Central to the analysis was a differential solubility fractionation (DSF) procedure using n-octyl-beta-D-glucopyranoside. The study achieved a significant increase (330) in the total number of known expressed proteins. From 612 identified, 185 were predicted to contain transmembrane domains or be associated with the membrane and 190 to be hydrophobic. The DSF procedure increased the efficacy of identifying membrane proteins by up to 169% and was economical, requiring far fewer runs (12% of machine time) to analyze the proteome compared to procedures without DSF. The analysis of peptide spectral counts enabled the assessment of growth temperature specific proteins. This semiquantitative analysis was particularly useful for identifying low abundance proteins unable to be quantified using labeling strategies. The proteogenomic analysis of the newly identified proteins revealed many cellular processes not previously associated with adaptation of the cell, This DSF-based approach is likely to benefit proteomic analyses of hydrophobic proteins for a broad range of biological systems.