Single-particle tracking (SPT) enables the ability to noninvasively probe the diffusive motions of individual proteins inside living cells at sub-diffraction-limit resolution. The stochastic motions of diffusing Rho GTPases encode information concerning its interactions with binding partners and with its local environment. By identifying Rho GTPases' diffusive states, insight can thus be gained into the spatiotemporal in vivo biochemistry inside live cells at a single-molecule resolution. Here we present perturbation expectation-maximization (pEM), a computational method which simultaneously analyzes a population of protein trajectories to uncover the system of diffusive behaviors: (1) the number of diffusive states, (2) the properties of each such diffusive state, and (3) the probabilities of each trajectory to a respective diffusive state. We provide a step-by-step guide to pEM and discuss considerations for its practical applications, including pEM's capabilities and limitations.
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http://dx.doi.org/10.1007/978-1-4939-8612-5_5 | DOI Listing |
PLoS Comput Biol
July 2024
Oden Institute, The University of Texas at Austin, Austin, Texas, United States of America.
The practical application of new single molecule protein sequencing (SMPS) technologies requires accurate estimates of their associated sequencing error rates. Here, we describe the development and application of two distinct parameter estimation methods for analyzing SMPS reads produced by fluorosequencing. A Hidden Markov Model (HMM) based approach, extends whatprot, where we previously used HMMs for SMPS peptide-read matching.
View Article and Find Full Text PDFbioRxiv
July 2023
Oden Institute, The University of Texas at Austin, Austin, TX 78712.
The practical application of new single molecule protein sequencing (SMPS) technologies requires accurate estimates of their associated sequencing error rates. Here, we describe the development and application of two distinct parameter estimation methods for analyzing SMPS reads produced by fluorosequencing. A Hidden Markov Model (HMM) based approach, extends , where we previously used HMMs for SMPS peptide-read matching.
View Article and Find Full Text PDFEur Phys J E Soft Matter
June 2023
Integrated Graduate Program in Physical and Engineering Biology, Yale University, New Haven, CT, 06511, USA.
We present and analyze video-microscopy-based single-particle-tracking measurements of the budding yeast (Saccharomyces cerevisiae) membrane protein, Pma1, fluorescently labeled either by direct fusion to the switchable fluorescent protein, mEos3.2, or by a novel, light-touch, labeling scheme, in which a 5 amino acid tag is directly fused to the C-terminus of Pma1, which then binds mEos3.2.
View Article and Find Full Text PDFJ Appl Stat
April 2023
Faculty of Science and Technology, University of Canberra, Canberra, Australia.
PLoS One
December 2022
Graduate School of Information, Yonsei University, Seoul, Korea.
Recent advances in positioning techniques, along with the widespread use of mobile devices, make it easier to monitor and collect user trajectory information during their daily activities. An ever-growing abundance of data about trajectories of individual users paves the way for various applications that utilize user mobility information. One of the most common analysis tasks in these new applications is to extract the sequential transition patterns between two consecutive timestamps from a collection of trajectories.
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