Automated Assignment of Proteoform Classification Levels.

Automated Assignment of Proteoform Classification Levels.
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DOI:
10.1021/acs.jproteome.1c00417
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发表时间:
2021-08-06
影响因子:
4.4
通讯作者:
Smith LM
Smith LM
中科院分区:
生物学2区
文献类型:
--
作者:
Rolfs Z;Smith LM

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蛋白质形式鉴定是完全了解样本中存在的生物多样性所必需的。然而,这些鉴定往往是模棱两可的,因为在用质谱学分析全长蛋白质方面存在挑战。最近开发了一个五级蛋白质形式分类系统,以描述蛋白质形式识别的模糊性,并允许跨软件平台和获取方法进行比较。该系统的广泛采用需要软件工具来提供蛋白质形式鉴定的分类。这里我们描述了在软件程序MetaMorpheus中实现的五级分类系统,它提供自下而上和自上而下的标识。此外,我们还开发了一个名为Proteoform分类器的独立程序,它允许用户对来自任何搜索程序的蛋白质形式结果进行分类,前提是该程序编写的输出包括评估蛋白质形式歧义所需的信息。这个独立的程序包括一个小的测试文件和数据库,用来评估给定程序是否提供了足够的信息来评估歧义。如果程序没有这样做,则Proteoform分类器将提供有意义的反馈,以帮助开发人员实现分类系统。我们测试了目前可用的自上而下的软件程序,发现除了MetaMorpheus之外,没有一个程序提供了关于身份模糊的足够信息,从而可以进行分类。
Proteoform identification is required to fully understand the biological diversity present in a sample. However, these identifications are often ambiguous because of the challenges in analyzing full length proteins by mass spectrometry. A five-level proteoform classification system was recently developed to delineate the ambiguity of proteoform identifications and to allow for comparisons across software platforms and acquisition methods. Widespread adoption of this system requires software tools to provide classification of the proteoform identifications. We describe here implementation of the five-level classification system in the software program MetaMorpheus, which provides both bottom-up and top-down identifications. Additionally, we developed a stand-alone program called ProteoformClassifier that allows users to classify proteoform results from any search program, provided that the program writes output that includes the information necessary to evaluate proteoform ambiguity. This stand-alone program includes a small test file and database to evaluate if a given program provides sufficient information to evaluate ambiguity. If the program does not, then ProteoformClassifier provides meaningful feedback to assist developers with implementing the classification system. We tested currently available top-down software programs and found that none of them other than MetaMorpheus provided sufficient information regarding identification ambiguity to permit classification.
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