Whiteaker Jeffrey R, Warscheid Bettina, Pribil Partick, Hathout Yetrib, Fenselau Catherine
Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
J Mass Spectrom. 2004 Oct;39(10):1113-21. doi: 10.1002/jms.668.
Three abundant small acid-soluble proteins (SASPs) from spores of Bacillus globigii were sequenced using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with post-source decay and nanoelectrospray collision-induced dissociation tandem mass spectrometry. The proteins were extracted from spores with 1 M HCl. Scanning electron micrographs of spores before and after acid extraction show that the spores retain their overall structure but have a shriveled texture following the acid treatment. Extracted SASPs were purified by high-performance liquid chromatography and molecular masses of the SASPs were identified at 7068 (SASP-1), 7332 (SASP-2), and 8889 (gamma-SASP). De novo peptide sequencing was used to determine the protein sequences. The correct ordering of peptide sequences was aided by mapping overlapping enzymatic digests and by comparison with homologous SASPs from Bacillus stearothermophilus. B. globigii is used in many field tests as a surrogate for B. anthracis. Thus complete SASP sequences from B. globigii will facilitate the development of methods for rapid identification of bacteria based on mass spectrometry and the examination of taxonomic relationships between Bacillus species.
利用带有源后衰变的基质辅助激光解吸/电离飞行时间质谱和纳米电喷雾碰撞诱导解离串联质谱，对来自球状芽孢杆菌孢子的三种丰富的小酸溶性蛋白（SASP）进行了测序。这些蛋白用1 M盐酸从孢子中提取。酸提取前后孢子的扫描电子显微镜图像显示，孢子保留了其整体结构，但酸处理后质地皱缩。提取的SASP通过高效液相色谱法进行纯化，鉴定出SASP的分子量分别为7068（SASP-1）、7332（SASP-2）和8889（γ-SASP）。采用从头肽测序法确定蛋白质序列。通过绘制重叠酶切图谱以及与嗜热脂肪芽孢杆菌的同源SASP进行比较，辅助确定肽序列的正确排序。球状芽孢杆菌在许多现场试验中用作炭疽芽孢杆菌的替代物。因此，来自球状芽孢杆菌的完整SASP序列将有助于基于质谱的细菌快速鉴定方法的开发以及芽孢杆菌属物种间分类关系的研究。
