Elucidating the Stoichiometries of Host–Pathogen Protein Interactions with Mass Photometry

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Bacterial Pathogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2674))

Abstract

Mass photometry (MP) is a single molecule technique that enables the characterization of individual proteins. Here we show a detailed workflow using the Refeyn OneMP to investigate molecular complexes, using the M53 protein, a plasminogen-binding group A streptococcal M-like protein (PAM), and human plasminogen as exemplar proteins. The methodology described herein confirmed a 1:1 binding stoichiometry for the M53–plasminogen complex. Additionally, MP was used to identify the oligomerization state, homogeneity, purity, and approximate molecular weights of each protein.

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Correspondence to Martina Sanderson-Smith .

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Proctor, EJ., Satapathy, S., Sanderson-Smith, M. (2023). Elucidating the Stoichiometries of Host–Pathogen Protein Interactions with Mass Photometry. In: Nordenfelt, P., Collin, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology, vol 2674. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3243-7_13

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  • DOI: https://doi.org/10.1007/978-1-0716-3243-7_13

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3242-0

  • Online ISBN: 978-1-0716-3243-7

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