Single-molecule tracking for studying protein dynamics and target-search mechanism in live cells of S. cerevisiae

Single-molecule tracking (SMT) is a powerful approach to quantify the biophysical parameters of protein dynamics in live cells. Here, we describe a protocol for SMT in live cells of the budding yeast Saccharomyces cerevisiae. We detail how to genetically engineer yeast strains for SMT, how to set up...

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Detalles Bibliográficos
Autores principales: Podh, Nitesh Kumar, Das, Ayan, Dey, Partha, Paliwal, Sheetal, Mehta, Gunjan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9732415/
https://www.ncbi.nlm.nih.gov/pubmed/36595957
http://dx.doi.org/10.1016/j.xpro.2022.101900
Descripción
Sumario:Single-molecule tracking (SMT) is a powerful approach to quantify the biophysical parameters of protein dynamics in live cells. Here, we describe a protocol for SMT in live cells of the budding yeast Saccharomyces cerevisiae. We detail how to genetically engineer yeast strains for SMT, how to set up image acquisition parameters, and how different software programs can be used to quantify a variety of biophysical parameters such as diffusion coefficient, residence time, bound fraction, jump angles, and target-search parameters. For complete details on the use and execution of this protocol, please refer to Mehta et al. (1) and Ball et al..(2)

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