Speciation of coagulase negative staphylococci, isolated from indoor air, using SDS page gel bands of expressed proteins followed by MALDI TOF MS and MALDI TOF-TOF MS-MS analysis of tryptic peptides
Speciation of coagulase negative staphylococci, isolated from indoor air, using SDS page gel bands of expressed proteins followed by MALDI TOF MS and MALDI TOF-TOF MS-MS analysis of tryptic peptides
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DOI：
10.1016/j.mimet.2010.12.007
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发表时间：
2011-02-01
影响因子：
2.2
通讯作者：
Walla, Michael
中科院分区：
文献类型：
--
作者：
Fox, Karen;Fox, Alvin;Walla, Michael
本研究以一群从学校居室内空气中分离的凝固酶阴性葡萄球菌为研究对象。细胞提取物和生理学试验（包括API STAPH）的常规MALDI TOF MS分析提供了该组菌株的不完整鉴定。使用1D凝胶电泳分离100 kDa条带后，使用MALDI TOF MS分析肽（胰蛋白酶消化释放），除了确定来源蛋白（乌头酸水合酶）的身份外，还可以改善细菌物种形成。这是通过Mascot搜索，经验观察和计算机生成的交叉相关分析的环境菌株与参考菌株。研究的物种包括一些基因组测序和其他未测序的基因组。使用MALDI TOF-TOF MS-MS分析代表可变和保守序列的一组不同的m/z值，证实肽序列来源于乌头酸水合酶。这里所描述的方法可能有广泛的应用在不同来源的环境分离物的物种形成和鉴定其表达的蛋白质。(C)2010 Elsevier B. V.保留所有权利。
A group of coagulase negative staphylococcal strains isolated from indoor air of occupied school rooms were the subject of this study. Conventional MALDI TOF MS profiling of cellular extracts and physiological tests (including API STAPH) provided incomplete identification of the set of strains. After separation of a 100 kDa band using 1D gel electrophoresis, profiling of peptides (released with tryptic digestion) using MALDI TOF MS allowed improved bacterial speciation in addition to determination of the identity of the protein of origin (aconitate hydratase). This was performed by Mascot search, empirical observation and computer-generated cross-correlation analysis of environmental isolates versus reference strains. The species studied included some with sequenced genomes and others with un-sequenced genomes. Peptide sequences were confirmed to originate from aconitate hydratase using MALDI TOF-TOF MS-MS analysis of a diverse set of m/z values representing variable and conserved sequences. The methodological approach described here might have widespread application in speciation of environmental isolates of diverse origin and in identification of their expressed proteins. (C) 2010 Elsevier B.V. All rights reserved.