Dworzanski Jacek P, Snyder A Peter
Science Applications International Corporation (SAIC), PO Box 68, Aberdeen Proving Ground, MD 21010-0068, USA.
Expert Rev Proteomics. 2005 Dec;2(6):863-78. doi: 10.1586/14789450.2.6.863.
Timely classification and identification of bacteria is of vital importance in many areas of public health. Mass spectrometry-based methods provide an attractive alternative to well-established microbiologic procedures. Mass spectrometry methods can be characterized by the relatively high speed of acquiring taxonomically relevant information. Gel-free mass spectrometry proteomics techniques allow for rapid fingerprinting of bacterial proteins using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry or, for high-throughput sequencing of peptides from protease-digested cellular proteins, using mass analysis of fragments from collision-induced dissociation of peptide ions. The latter technique uses database searching of product ion mass spectra. A database contains a comprehensive list of protein sequences translated from protein-encoding open reading frames found in bacterial genomes. The results of such searches allow the assignment of experimental peptide sequences to matching theoretical bacterial proteomes. Phylogenetic profiles of sequenced peptides are then used to create a matrix of sequence-to-bacterium assignments, which are analyzed using numerical taxonomy tools. The results thereof reveal the relatedness between bacteria, and allow the taxonomic position of an investigated strain to be inferred.
及时对细菌进行分类和鉴定在公共卫生的许多领域都至关重要。基于质谱的方法为成熟的微生物学程序提供了一种有吸引力的替代方案。质谱方法的特点是获取分类学相关信息的速度相对较快。无凝胶质谱蛋白质组学技术可使用基质辅助激光解吸/电离飞行时间质谱对细菌蛋白质进行快速指纹识别，或者对于蛋白酶消化的细胞蛋白质肽段的高通量测序，可使用肽离子碰撞诱导解离碎片的质量分析。后一种技术使用产物离子质谱的数据库搜索。数据库包含从细菌基因组中发现的蛋白质编码开放阅读框翻译而来的蛋白质序列的综合列表。此类搜索结果可将实验肽序列与匹配的理论细菌蛋白质组进行比对。然后，测序肽段的系统发育图谱用于创建序列与细菌比对的矩阵，并使用数值分类学工具进行分析。其结果揭示了细菌之间的亲缘关系，并可推断出被研究菌株的分类地位。
