Hands-On Electrospray Ionization Mass Spectrometry for Undergraduate Biochemistry Students: Peptide Identification by Ladder Sequencing,Journal of Chemical Education

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Hands-On Electrospray Ionization Mass Spectrometry for Undergraduate Biochemistry Students: Peptide Identification by Ladder Sequencing
Journal of Chemical Education ( IF 2.5 ) Pub Date : 2020-03-26 , DOI: 10.1021/acs.jchemed.9b00800
Viveka Perera ₁ , Baku Acharya ₁ , Amanda L. Patrick ₁ , Deb Mlsna ₁

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Mass spectrometers are ever-increasingly powerful, user-friendly, and affordable. Thus, the addition of mass spectrometry experiments into the undergraduate laboratory curriculum is now both feasible and an effective tool to introduce students to relevant instrumentation. Here an experiment demonstrating the use of a high-resolution electrospray ionization mass spectrometer (ESI-HRMS) for the identification of peptide sequences and the differentiation of isomeric peptides was developed and implemented in a biochemistry laboratory course for third-year chemistry major undergraduate students. This lab activity was a first introduction to mass spectrometry for undergraduate biochemistry students. As a prelab assignment, students were tasked to predict the mass-to-charge ratios and the fragmentation patterns for six given peptides using an online fragmentation prediction tool (Protein Prospector’s MS-Product tool). Students then analyzed two unknown peptides using ESI-HRMS. The theoretical and experimental results were compared to reveal the identity of the two unknown peptides. The student success rate of recognizing the unknown peptide sequences was 87.5%. This laboratory experiment provided students with hands-on experience using a research-grade ESI-HRMS instrument to solve a bioanalytical problem—specifically, the identification of the primary structure of a peptide of unknown sequence. Learning outcomes were evaluated for this experiment, which showed student understanding of peptide sequencing using mass spectrometry.

中文翻译：

面向生物化学本科生的动手电喷雾电离质谱：梯式测序鉴定肽

质谱仪功能日益强大，用户友好且价格合理。因此，将质谱实验添加到本科实验室课程中既可行，又是将学生介绍给相关仪器的有效工具。在此，针对三年级化学专业的本科生，在生物化学实验室课程中开发并实施了一个实验，该实验证明了使用高分辨率电喷雾电离质谱仪（ESI-HRMS）来鉴定肽序列和区分异构肽。该实验室活动是针对生物化学本科生的质谱学的首次介绍。作为预备作业，学生需要使用在线断裂预测工具（Protein Prospector的MS-Product工具）来预测六种给定肽的质荷比和断裂模式。然后，学生使用ESI-HRMS分析了两种未知肽。比较了理论和实验结果以揭示两种未知肽的身份。学生识别未知肽序列的成功率为87.5％。这项实验室实验为学生提供了使用研究级ESI-HRMS仪器解决生物分析问题的实践经验，特别是鉴定未知序列肽的一级结构。本实验评估了学习成果，表明学生对使用质谱法进行肽测序的理解。

更新日期：2020-03-26

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