Rapid chemical digestion of small acid-soluble spore proteins for analysis of Bacillus spores
Rapid chemical digestion of small acid-soluble spore proteins for analysis of Bacillus spores
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DOI：
10.1021/ac051521d
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发表时间：
2006-01-01
影响因子：
7.4
通讯作者：
Fenselau, C
中科院分区：
文献类型：
--
作者：
Swatkoski, S;Russell, SC;Fenselau, C
提出了一种基于小酸溶性孢子蛋白（SASP）的选择性释放和化学消化的芽孢杆菌孢子的快速鉴定方法。微波辅助酸水解炭疽杆菌中的 SASP。斯特恩和枯草芽孢杆菌 str。 168 是一步完成的，仅需要 90 秒的加热。使用配备弯曲场反射器的 MALDI-TOF-MS 通过源后衰变测序来鉴定化学消化的肽产物。使用跨物种序列搜索评估观察到的 SASP 肽的特异性。在这些条件下酸消化的不完全性质允许检测消化产物及其来源的蛋白质，这增加了物种鉴定的可信度。通过分析枯草芽孢杆菌的混合物，进一步证明了这种快速鉴定芽孢杆菌属的方法的可行性。 168 和 B. anthracis str.斯特恩孢子。
A method for the rapid identification of Bacillus spores is proposed, based on the selective release and chemical digestion of small, acid-soluble spore proteins (SASPs). Microwave-assisted acid hydrolysis of SASPs from B. anthracis str. Sterne and B. subtilis str. 168 was accomplished in a single step requiring only 90 s of heating. The peptide products of the chemical digestion were identified by postsource decay sequencing with a MALDI-TOF-MS equipped with a curved-field reflectron. The specificity of the observed SASP peptides was evaluated using a cross-species sequence search. The incomplete nature of the acid digestion under these conditions allowed detection of the digest products along with the proteins from which they originated, which increased species identification confidence. The feasibility of this approach for the rapid identification of Bacillus species was further demonstrated by analyzing a mixture of B. subtilis str. 168 and B. anthracis str. Sterne spores.