Diffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells

Single-particle tracking enables the analysis of the dynamics of biomolecules in living cells with nanometer spatial and millisecond temporal resolution. This technique reports on the mobility of membrane proteins and is sensitive to the molecular state of a biomolecule and to interactions with othe...

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Autores principales:	Johanna V. Rahm, Sebastian Malkusch, Ulrike Endesfelder, Marina S. Dietz, Mike Heilemann
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	single-particle tracking single-trajectory analysis membrane receptors MET receptor single-molecule imaging diffusion states Electronic computers. Computer science QA75.5-76.95
Acceso en línea:	https://doaj.org/article/857c69d2d420412682edc4651897ba0b
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id	oai:doaj.org-article:857c69d2d420412682edc4651897ba0b
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spelling	oai:doaj.org-article:857c69d2d420412682edc4651897ba0b2021-11-12T06:04:54ZDiffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells2624-989810.3389/fcomp.2021.757653https://doaj.org/article/857c69d2d420412682edc4651897ba0b2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fcomp.2021.757653/fullhttps://doaj.org/toc/2624-9898Single-particle tracking enables the analysis of the dynamics of biomolecules in living cells with nanometer spatial and millisecond temporal resolution. This technique reports on the mobility of membrane proteins and is sensitive to the molecular state of a biomolecule and to interactions with other biomolecules. Trajectories describe the mobility of single particles over time and provide information such as the diffusion coefficient and diffusion state. Changes in particle dynamics within single trajectories lead to segmentation, which allows to extract information on transitions of functional states of a biomolecule. Here, mean-squared displacement analysis is developed to classify trajectory segments into immobile, confined diffusing, and freely diffusing states, and to extract the occurrence of transitions between these modes. We applied this analysis to single-particle tracking data of the membrane receptor MET in live cells and analyzed state transitions in single trajectories of the un-activated receptor and the receptor bound to the ligand internalin B. We found that internalin B-bound MET shows an enhancement of transitions from freely and confined diffusing states into the immobile state as compared to un-activated MET. Confined diffusion acts as an intermediate state between immobile and free, as this state is most likely to change the diffusion state in the following segment. This analysis can be readily applied to single-particle tracking data of other membrane receptors and intracellular proteins under various conditions and contribute to the understanding of molecular states and signaling pathways.Johanna V. RahmSebastian MalkuschUlrike EndesfelderUlrike EndesfelderMarina S. DietzMike HeilemannFrontiers Media S.A.articlesingle-particle trackingsingle-trajectory analysismembrane receptorsMET receptorsingle-molecule imagingdiffusion statesElectronic computers. Computer scienceQA75.5-76.95ENFrontiers in Computer Science, Vol 3 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	single-particle tracking single-trajectory analysis membrane receptors MET receptor single-molecule imaging diffusion states Electronic computers. Computer science QA75.5-76.95
spellingShingle	single-particle tracking single-trajectory analysis membrane receptors MET receptor single-molecule imaging diffusion states Electronic computers. Computer science QA75.5-76.95 Johanna V. Rahm Sebastian Malkusch Ulrike Endesfelder Ulrike Endesfelder Marina S. Dietz Mike Heilemann Diffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells
description	Single-particle tracking enables the analysis of the dynamics of biomolecules in living cells with nanometer spatial and millisecond temporal resolution. This technique reports on the mobility of membrane proteins and is sensitive to the molecular state of a biomolecule and to interactions with other biomolecules. Trajectories describe the mobility of single particles over time and provide information such as the diffusion coefficient and diffusion state. Changes in particle dynamics within single trajectories lead to segmentation, which allows to extract information on transitions of functional states of a biomolecule. Here, mean-squared displacement analysis is developed to classify trajectory segments into immobile, confined diffusing, and freely diffusing states, and to extract the occurrence of transitions between these modes. We applied this analysis to single-particle tracking data of the membrane receptor MET in live cells and analyzed state transitions in single trajectories of the un-activated receptor and the receptor bound to the ligand internalin B. We found that internalin B-bound MET shows an enhancement of transitions from freely and confined diffusing states into the immobile state as compared to un-activated MET. Confined diffusion acts as an intermediate state between immobile and free, as this state is most likely to change the diffusion state in the following segment. This analysis can be readily applied to single-particle tracking data of other membrane receptors and intracellular proteins under various conditions and contribute to the understanding of molecular states and signaling pathways.
format	article
author	Johanna V. Rahm Sebastian Malkusch Ulrike Endesfelder Ulrike Endesfelder Marina S. Dietz Mike Heilemann
author_facet	Johanna V. Rahm Sebastian Malkusch Ulrike Endesfelder Ulrike Endesfelder Marina S. Dietz Mike Heilemann
author_sort	Johanna V. Rahm
title	Diffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells
title_short	Diffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells
title_full	Diffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells
title_fullStr	Diffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells
title_full_unstemmed	Diffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells
title_sort	diffusion state transitions in single-particle trajectories of met receptor tyrosine kinase measured in live cells
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/857c69d2d420412682edc4651897ba0b
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Diffusion State Transitions in Single-Particle Trajectories of MET Receptor Tyrosine Kinase Measured in Live Cells

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