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ACS Appl Mater Interfaces. 2017 Aug 30;9(34):28168-28179. doi: 10.1021/acsami.7b04622. Epub 2017 Aug 21.

Parallel Compression Is a Fast Low-Cost Assay for the High-Throughput Screening of Mechanosensory Cytoskeletal Proteins in Cells.

Author information

1
CAS Key Laboratory of Mechanical Behavior and Design of Materials, Department of Modern Mechanics, University of Science and Technology of China , Hefei 230000, China.
2
Departments of Cell Biology, Pharmacology and Molecular Medicine, and Medicine, School of Medicine, Johns Hopkins University , Baltimore, Maryland 21205, United States.
3
Department of Chemical and Biomolecular Engineering, Whiting School of Engineering, Johns Hopkins University , Baltimore, Maryland 21211, United States.

Abstract

Cellular mechanosensing is critical for many biological processes, including cell differentiation, proliferation, migration, and tissue morphogenesis. The actin cytoskeletal proteins play important roles in cellular mechanosensing. Many techniques have been used to investigate the mechanosensory behaviors of these proteins. However, a fast, low-cost assay for the quantitative characterization of these proteins is still lacking. Here, we demonstrate that compression assay using agarose overlay is suitable for the high throughput screening of mechanosensory proteins in live cells while requiring minimal experimental setup. We used several well-studied myosin II mutants to assess the compression assay. On the basis of elasticity theories, we simulated the mechanosensory accumulation of myosin II's and quantitatively reproduced the experimentally observed protein dynamics. Combining the compression assay with confocal microscopy, we monitored the polarization of myosin II oligomers at the subcellular level. The polarization was dependent on the ratio of the two principal strains of the cellular deformations. Finally, we demonstrated that this technique could be used on the investigation of other mechanosensory proteins.

KEYWORDS:

actin filament; compression; mechanobiology; mechanosensing; mechanosensory accumulation; mechanotransduction; myosin II

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