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Species Differentiation of a Diverse Suite of Bacillus Spores by Mass Spectrometry-Based Protein Profiling

机译：基于质谱的蛋白质谱分析芽孢杆菌孢子的不同套件的物种分化

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摘要

In this study, we demonstrate the versatility of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOFMS) protein profiling for the species differentiation of a diverse suite of Bacillus spores. MALDI-TOFMS protein profiles of 11 different strains of Bacillus spores, encompassing nine different species, were evaluated. Bacillus species selected for MALDI-TOFMS analysis represented the spore-forming bacterial diversity of typical class 100K clean room spacecraft assembly facilities. A one-step sample treatment and MALDI-TOFMS preparation were used to minimize the sample preparation time. A library of MALDI-TOFMS spectra was created from these nine Bacillus species, the most diverse protein profiling study of the genus reported to date. Linear correlation analysis was used to successfully differentiate the MALDI-TOFMS protein profiles from all strains evaluated in this study. The MALDI-TOFMS protein profiles were compared with 16S rDNA sequences for their bacterial systematics and molecular phylogenetic affiliations. The MALDI-TOFMS profiles were found to be complementary to the 16S rDNA analysis. Proteomic studies of Bacillus subtilis 168 were pursued to identify proteins represented by the biomarker peaks in the MALDI-TOFMS spectrum. Four small, acid-soluble proteins (A, B, C, and D), one DNA binding protein, hypothetical protein ymf J, and four proteins associated with the spore coat and spore coat formation (coat JB, coat F, coat T, and spoIVA) were identified. The ability to visualize higher-molecular-mass coat proteins (10 to 25 kDa) as well as smaller proteins (<10 kDa) with MALDI-TOFMS profiling is critical for the complete and effective species differentiation of the Bacillus genus.
机译：在这项研究中，我们证明了基质辅助激光解吸电离飞行时间质谱（MALDI-TOFMS）蛋白谱的多样性，可用于多种芽孢杆菌孢子的物种分化。评估了11种不同菌株的芽孢杆菌MALDI-TOFMS蛋白质谱，这些菌株涵盖9种不同菌种。选择用于MALDI-TOFMS分析的芽孢杆菌物种代表了典型的100K级洁净室航天器组装设施中形成孢子的细菌多样性。一站式样品处理和MALDI-TOFMS制备可最大程度地减少样品制备时间。从这九种芽孢杆菌属物种创建了MALDI-TOFMS光谱库，这是迄今为止报道的该属中最多样化的蛋白质谱研究。线性相关分析用于成功地区分本研究中评估的所有菌株的MALDI-TOFMS蛋白谱。将MALDI-TOFMS蛋白质谱与16S rDNA序列进行细菌系统学和分子系统发育关系的比较。发现MALDI-TOFMS配置文件与16S rDNA分析互补。进行了枯草芽孢杆菌168的蛋白质组学研究，以鉴定MALDI-TOFMS谱中生物标志物峰代表的蛋白质。四种小的酸溶蛋白（A，B，C和D），一种DNA结合蛋白，假定蛋白ymf J和四种与孢子外壳和孢子外壳形成相关的蛋白（外壳JB，外壳F，外壳T，和spoIVA）。 MALDI-TOFMS分析显示较高分子量的外壳蛋白（10至25 kDa）和较小的蛋白（<10 kDa）的能力对于芽孢杆菌属的完整有效的物种分化至关重要。

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