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完整蛋白质复合物的化学交联和质谱分析研究多亚基蛋白质组合体的体系结构

JoVE Journal
Biology

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JoVE Journal Biology

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

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完整蛋白质复合物的化学交联和质谱分析研究多亚基蛋白质组合体的体系结构

DOI:

10.3791/56747-v

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10:01 min

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November 28, 2017

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Caroline Haupt*¹, Tommy Hofmann*¹, Sabine Wittig*¹, Susann Kostmann, Argyris Politis, Carla Schmidt

¹Interdisciplinary research center HALOmem,Martin Luther University Halle-Wittenberg, ²Department of Chemistry,Kings College London

Chapters

00:05Title
00:51Mass Spectrometry-based Protein Identification
04:35Mass Spectrometry of Intact Protein Complexes (Native Mass Spectrometry)
06:17Chemical Cross-linking Coupled with Mass Spectrometry
07:19Results: Chemical Cross-linking and Mass Spectrometry of Multi-subunit Protein Complexes
09:14Conclusion

Summary

Automatic Translation

蛋白质复合体的结构对其功能至关重要。结合各种质谱技术证明了研究其组装的强大功能。我们提供化学交联和本机质谱的协议, 并展示这些互补技术如何帮助阐明 multi-subunit 蛋白组件的结构。

Tags

Chemical Cross-linking Mass Spectrometry Intact Protein Complexes Multi-subunit Protein Assemblies Structural Information Composition Connectivity Stoichiometry Protein Complex Purification Electrophoresis Coomassie Staining Protein Band Excision Disulfide Reduction Cysteine Alkylation Trypsin Digestion Peptide Extraction

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