液体常压MALDI质谱法分析真菌的物种形成。

IF 3.8 2区 化学 Q1 BIOCHEMICAL RESEARCH METHODS
Lily R Adair, Ian M Jones, Rainer Cramer
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引用次数: 0

摘要

真菌病原体对全球健康构成越来越大的威胁,需要快速准确的鉴定方法。在这里,液体常压基质辅助激光解吸/电离(LAP-MALDI)质谱(MS)被应用于白色念珠菌和酿酒酵母培养菌落的快速脂质和蛋白质谱分析。在m/z 600-1100范围内观察到物种特异性脂质谱,经串联质谱(MS/MS)证实,以磷脂为主。经过简单的固相萃取清理,LAP-MALDI质谱显示了适合MS/MS分析的多种带电蛋白离子。对于白色假丝酵母菌,完全成熟的物种特异性WHS11蛋白(~ 7 kDa; P43074)被完整地检测到,并通过自上而下的MS/MS蛋白形态测序,包括n端甲硫氨酸引发物的切割和相关的n端乙酰化,得到了可靠的鉴定。对于酿酒酵母,采用MS/MS分析测序了一组蛋白质形态,并在“UniProtKB参考蛋白质组+ Swiss-Prot”目标数据库中鉴定了两个物种特异性蛋白质。其中一个也被完整地检测到,并被测序和鉴定为完全成熟的HSP12蛋白(~ 11.5 kDa; P22943)。这项工作证明了LAP-MALDI MS和MS/MS生物分型作为一种强大的、无标签的快速真菌分类和蛋白质形态表征平台的潜力,比传统的MALDI生物分型有了实质性的改进。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Speciation analysis of fungi by liquid atmospheric pressure MALDI mass spectrometry.

Fungal pathogens pose a growing threat to global health, necessitating rapid and accurate identification methods. Here, liquid atmospheric pressure matrix-assisted laser desorption/ionisation (LAP-MALDI) mass spectrometry (MS) is applied to fast lipid and protein profiling of Candida albicans and Saccharomyces cerevisiae from cultured colonies. Species-specific lipid profiles were observed in the m/z 600-1100 range, dominated by phospholipids as confirmed by tandem mass spectrometry (MS/MS). Following simple solid phase extraction clean-up, LAP-MALDI mass spectra revealed multiply charged protein ions suitable for MS/MS analysis. For C. albicans, the fully mature, species-specific WHS11 protein (~ 7 kDa; P43074) was detected intact and confidently identified by top-down MS/MS proteoform sequencing, including the cleavage of the N-terminal methionine initiator and the associated N-terminal acetylation. For S. cerevisiae, a set of proteoforms were sequenced by MS/MS analysis, which led to the identification of two species-specific proteins within the 'UniProtKB reference proteomes + Swiss-Prot' target database. One of these was also detected intact, and sequenced and identified as the fully mature HSP12 protein (~ 11.5 kDa; P22943). This work demonstrates the potential of LAP-MALDI MS and MS/MS biotyping as a powerful, label-free platform for rapid fungal classification and proteoform characterisation, offering substantial improvements over conventional MALDI biotyping.

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来源期刊
CiteScore
8.00
自引率
4.70%
发文量
638
审稿时长
2.1 months
期刊介绍: Analytical and Bioanalytical Chemistry’s mission is the rapid publication of excellent and high-impact research articles on fundamental and applied topics of analytical and bioanalytical measurement science. Its scope is broad, and ranges from novel measurement platforms and their characterization to multidisciplinary approaches that effectively address important scientific problems. The Editors encourage submissions presenting innovative analytical research in concept, instrumentation, methods, and/or applications, including: mass spectrometry, spectroscopy, and electroanalysis; advanced separations; analytical strategies in “-omics” and imaging, bioanalysis, and sampling; miniaturized devices, medical diagnostics, sensors; analytical characterization of nano- and biomaterials; chemometrics and advanced data analysis.
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