Manish Singh Kushwah

Label-free tracking and mass measurement of single proteins on lipid bilayers

Publisher: Cold Spring Harbor Laboratory

Date: 08-04-2021

DOI: 10.1101/2021.04.08.438951

Abstract: We introduce dynamic mass photometry, a method for label-free imaging, tracking and mass measurement of membrane-associated proteins. Our method enables quantitative studies of their mobility, membrane affinity and interactions at the single molecule level. Application to the membrane remodelling GTPase dynamin1 reveals heterogeneous mixtures of oligomers suggesting that the fundamental building block for oligomerisation is a dimer, challenging current tetramer-centric models. Dynamic mass photometry has the ability to transform our approach to studying biomolecular mechanisms in and on lipid bilayers.

Mass photometry enables label-free tracking and mass measurement of single proteins on lipid bilayers

Publisher: Springer Science and Business Media LLC

Date: 10-2021

DOI: 10.1038/S41592-021-01261-W

Abstract: The quantification of membrane-associated biomolecular interactions is crucial to our understanding of various cellular processes. State-of-the-art single-molecule approaches rely largely on the addition of fluorescent labels, which complicates the quantification of the involved stoichiometries and dynamics because of low temporal resolution and the inherent limitations associated with labeling efficiency, photoblinking and photobleaching. Here, we demonstrate dynamic mass photometry, a method for label-free imaging, tracking and mass measurement of in idual membrane-associated proteins diffusing on supported lipid bilayers. Application of this method to the membrane remodeling GTPase, dynamin-1, reveals heterogeneous mixtures of dimer-based oligomers, oligomer-dependent mobilities, membrane affinities and (dis)association of in idual complexes. These capabilities, together with assay-based advances for studying integral membrane proteins, will enable the elucidation of biomolecular mechanisms in and on lipid bilayers.

The Maharaja Sayajirao University Of Baroda Faculty Of Science

Location: India

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Indian Institute Of Science Education Research Pune

Location: India

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University Of Oxford

Location: United Kingdom of Great Britain and Northern Ireland

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