Identification of bacterial proteins observed in MALDI TOF mass spectra from whole cells
Identification of bacterial proteins observed in MALDI TOF mass spectra from whole cells
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DOI：
10.1021/ac990175v
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发表时间：
1999-08-01
影响因子：
7.4
通讯作者：
Lay, JO
中科院分区：
文献类型：
--
作者：
Holland, RD;Duffy, CR;Lay, JO
从细菌细胞的MALDI TOF质谱中的特征离子已与四个已知的蛋白质。通过HPLC分离从细胞和过滤的细胞悬浮液中观察到的蛋白质，并基于其质谱和使用Edman方法测定的其部分氨基酸序列（10-15个残基）进行鉴定。抗酸蛋白HdeA和HdeB产生来自细胞和来自大肠杆菌1090和福氏志贺菌PHS-1059两者的提取物的m/z 9735和9060 iu MALDI TOF质谱附近的离子。然而，与蛋白水解裂解的肽酶Lep，而不是前体蛋白，观察到的蛋白质，无论是使用细胞和细胞提取物。一种冷休克蛋白CspA与来自铜绿假单胞菌的m/z 7643附近的离子相关。类似地，冷适应蛋白CapB被鉴定为恶臭假单胞菌中m/z 7684附近的离子的来源，最后一种蛋白与来自脆弱假单胞菌的已知CapB同源。虽然这些实验涉及检测来自典型细菌的已知或同源蛋白质，但这种相同的方法也可以应用于检测与具有公共卫生意义的其他细菌相关的独特蛋白质或生物标志物蛋白质。
Characteristic ions in the MALDI TOF mass spectra from bacterial cells have been associated with four known proteins. The proteins, observed both from cells and in filtered cellular suspensions, were isolated by HPLC and identified on the basis of their mass spectra and their partial amino acid sequence, determined using the Edman method (10-15 residues). The acid resistance proteins HdeA and HdeB give rise to ions near m/z 9735 and 9060 iu MALDI TOF mass spectra from cells and from extracts of both Escherichia coli 1090 and Shigella flexneri PHS-1059. However, the proteins associated with proteolytic cleavage by the peptidase Lep, rather than the precursor proteins, were observed, both using cells and from cellular extracts. A cold-shock protein, CspA, was associated with the ion near m/z 7643 from Pseudomonas aeruginosa. Similarly, a cold-acclimation protein, CapB, was identified as the source of the ion near m/z 7684 in P. putida, This last protein was homologous with a known CapB from P. fragi. While these experiments involved the detection of known or homologous proteins from typical bacteria, this same approach could also be applied to the detection of unique proteins or biomarker proteins associated with other bacteria of public health significance.