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Visualizing the in vitro assembly of tropomyosin/actin filaments using TIRF microscopy.
Biophys Rev. 2020 Jul 07 [Online ahead of print]BR

Abstract

Tropomyosins are elongated alpha-helical proteins that form co-polymers with most actin filaments within a cell and play important roles in the structural and functional diversification of the actin cytoskeleton. How the assembly of tropomyosins along an actin filament is regulated and the kinetics of tropomyosin association with an actin filament is yet to be fully determined. A recent series of publications have used total internal reflection fluorescence (TIRF) microscopy in combination with advanced surface and protein chemistry to visualise the molecular assembly of actin/tropomyosin filaments in vitro. Here, we review the use of the in vitro TIRF assay in the determination of kinetic data on tropomyosin filament assembly. This sophisticated approach has enabled generation of real-time single-molecule data to fill the gap between in vitro bulk assays and in vivo assays of tropomyosin function. The in vitro TIRF assays provide a new foundation for future studies involving multiple actin-binding proteins that will more accurately reflect the physiological protein-protein interactions in cells.

Authors+Show Affiliations

School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia.School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia.School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia.School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia.School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia. p.gunning@unsw.edu.au.

Pub Type(s)

Journal Article
Review

Language

eng

PubMed ID

32638329

Citation

Janco, Miro, et al. "Visualizing the in Vitro Assembly of Tropomyosin/actin Filaments Using TIRF Microscopy." Biophysical Reviews, 2020.
Janco M, Dedova I, Bryce NS, et al. Visualizing the in vitro assembly of tropomyosin/actin filaments using TIRF microscopy. Biophys Rev. 2020.
Janco, M., Dedova, I., Bryce, N. S., Hardeman, E. C., & Gunning, P. W. (2020). Visualizing the in vitro assembly of tropomyosin/actin filaments using TIRF microscopy. Biophysical Reviews. https://doi.org/10.1007/s12551-020-00720-6
Janco M, et al. Visualizing the in Vitro Assembly of Tropomyosin/actin Filaments Using TIRF Microscopy. Biophys Rev. 2020 Jul 7; PubMed PMID: 32638329.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Visualizing the in vitro assembly of tropomyosin/actin filaments using TIRF microscopy. AU - Janco,Miro, AU - Dedova,Irina, AU - Bryce,Nicole S, AU - Hardeman,Edna C, AU - Gunning,Peter W, Y1 - 2020/07/07/ PY - 2020/05/31/received PY - 2020/07/02/accepted PY - 2020/7/9/entrez KW - Actin filaments KW - Actin-binding proteins KW - Intramolecular interactions KW - TIRF microscopy KW - Tropomyosin JF - Biophysical reviews JO - Biophys Rev N2 - Tropomyosins are elongated alpha-helical proteins that form co-polymers with most actin filaments within a cell and play important roles in the structural and functional diversification of the actin cytoskeleton. How the assembly of tropomyosins along an actin filament is regulated and the kinetics of tropomyosin association with an actin filament is yet to be fully determined. A recent series of publications have used total internal reflection fluorescence (TIRF) microscopy in combination with advanced surface and protein chemistry to visualise the molecular assembly of actin/tropomyosin filaments in vitro. Here, we review the use of the in vitro TIRF assay in the determination of kinetic data on tropomyosin filament assembly. This sophisticated approach has enabled generation of real-time single-molecule data to fill the gap between in vitro bulk assays and in vivo assays of tropomyosin function. The in vitro TIRF assays provide a new foundation for future studies involving multiple actin-binding proteins that will more accurately reflect the physiological protein-protein interactions in cells. SN - 1867-2450 UR - https://www.unboundmedicine.com/medline/citation/32638329/Visualizing_the_in_vitro_assembly_of_tropomyosin/actin_filaments_using_TIRF_microscopy L2 - https://dx.doi.org/10.1007/s12551-020-00720-6 DB - PRIME DP - Unbound Medicine ER -
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