Dissecting myosin-5B mechanosensitivity and calcium regulation at the single molecule level

Nature Communications
Lucia GardiniMarco Capitanio

Abstract

Myosin-5B is one of three members of the myosin-5 family of actin-based molecular motors. Despite its fundamental role in recycling endosome trafficking and in collective actin network dynamics, the molecular mechanisms underlying its motility are inherently unknown. Here we combine single-molecule imaging and high-speed laser tweezers to dissect the mechanoenzymatic properties of myosin-5B. We show that a single myosin-5B moves processively in 36-nm steps, stalls at ~2 pN resistive forces, and reverses its directionality at forces >2 pN. Interestingly, myosin-5B mechanosensitivity differs from that of myosin-5A, while it is strikingly similar to kinesin-1. In particular, myosin-5B run length is markedly and asymmetrically sensitive to force, a property that might be central to motor ensemble coordination. Furthermore, we show that Ca2+ does not affect the enzymatic activity of the motor unit, but abolishes myosin-5B processivity through calmodulin dissociation, providing important insights into the regulation of postsynaptic cargoes trafficking in neuronal cells.

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Citations

Dec 22, 2019·International Journal of Molecular Sciences·Natalia Fili, Christopher P Toseland
Sep 19, 2020·Physical Review. E·Hamid KhataeeZoltan Neufeld
Dec 15, 2019·Trends in Cell Biology·Sarah R BargerMira Krendel
Nov 24, 2021·Liver International : Official Journal of the International Association for the Study of the Liver·Li WangJian-She Wang

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Methods Mentioned

BETA
motility assay
optical tweezers
motility assays
co-immunoprecipitation
size exclusion chromatography
ion exchange chromatography
fluorescence microscopy

Software Mentioned

Fiji
Matlab
CellTrak ( Motion Analysis )
Labview

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