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

被引:10
|
作者
Rahm, Johanna V. [1 ]
Malkusch, Sebastian [2 ]
Endesfelder, Ulrike [3 ,4 ]
Dietz, Marina S. [1 ]
Heilemann, Mike [1 ]
机构
[1] Goethe Univ, Inst Phys & Theoret Chem, Frankfurt, Germany
[2] Goethe Univ, Inst Clin Pharmacol, Frankfurt, Germany
[3] Carnegie Mellon Univ, Dept Phys, Pittsburgh, PA 15213 USA
[4] Rhein Friedrich Wilhelms Univ Bonn, Inst Microbiol & Biotechnol, Bonn, Germany
来源
关键词
single-particle tracking; single-trajectory analysis; membrane receptors; MET receptor; single-molecule imaging; diffusion states; LISTERIA-MONOCYTOGENES; GROWTH-FACTOR; PROTEIN; TRACKING; DIMERIZATION; DYNAMICS; INLB; ACTIVATION; EXPRESSION; ENTRY;
D O I
10.3389/fcomp.2021.757653
中图分类号
TP39 [计算机的应用];
学科分类号
081203 ; 0835 ;
摘要
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.
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页数:8
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