TY - JOUR
T1 - Changes in single-molecule integrin dynamics linked to local cellular behavior
AU - Jaqaman, Khuloud
AU - Galbraith, James A.
AU - Davidson, Michael W.
AU - Galbraith, Catherine G.
N1 - Funding Information:
We thank H. Elliott and G. Danuser for cell mask and windowing software, J. R. Galbraith for circuit assistance, and M. Ginsberg and D. Calderwood for constructs. u-Track is available at www .utsouthwestern.edu/labs/jaqaman/software. Funding was provided by the National Institutes of Health Intramural Director's Challenge Award (C.G.G. and J.A.G.) and Super-resolution Initiative (C.G.G.), the OHSU Knight Cancer Institute (C.G.G. and J.A.G.), the intramural program of the National Institute of Neurological Disorders and Stroke/National Institutes of Health (J.A.G.), National Institutes of Health Grant P50 GM068762 (K.J.; P. Sorger, PI), and the UT Southwestern Endowed Scholars Program (K.J.).
Publisher Copyright:
© 2016 Jaqaman et al.
PY - 2016/5/15
Y1 - 2016/5/15
N2 - Recent advances in light microscopy permit visualization of the behavior of individual molecules within dense macromolecular ensembles in live cells. It is now conceptually possible to relate the dynamic organization of molecular machinery to cellular function. However, inherent heterogeneities, as well as disparities between spatial and temporal scales, pose substantial challenges in deriving such a relationship. New approaches are required to link discrete single-molecule behavior with continuous cellular-level processes. Here we combined intercalated molecular and cellular imaging with a computational framework to detect reproducible transient changes in the behavior of individual molecules that are linked to cellular behaviors. Applying our approach to integrin transmembrane receptors revealed a spatial density gradient underlying characteristic molecular density increases and mobility decreases, indicating the subsequent onset of local protrusive activity. Integrin mutants further revealed that these density and mobility transients are separable and depend on different binding domains within the integrin cytoplasmic tail. Our approach provides a generalizable paradigm for dissecting dynamic spatiotemporal molecular behaviors linked to local cellular events.
AB - Recent advances in light microscopy permit visualization of the behavior of individual molecules within dense macromolecular ensembles in live cells. It is now conceptually possible to relate the dynamic organization of molecular machinery to cellular function. However, inherent heterogeneities, as well as disparities between spatial and temporal scales, pose substantial challenges in deriving such a relationship. New approaches are required to link discrete single-molecule behavior with continuous cellular-level processes. Here we combined intercalated molecular and cellular imaging with a computational framework to detect reproducible transient changes in the behavior of individual molecules that are linked to cellular behaviors. Applying our approach to integrin transmembrane receptors revealed a spatial density gradient underlying characteristic molecular density increases and mobility decreases, indicating the subsequent onset of local protrusive activity. Integrin mutants further revealed that these density and mobility transients are separable and depend on different binding domains within the integrin cytoplasmic tail. Our approach provides a generalizable paradigm for dissecting dynamic spatiotemporal molecular behaviors linked to local cellular events.
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U2 - 10.1091/mbc.E16-01-0018
DO - 10.1091/mbc.E16-01-0018
M3 - Article
C2 - 27009207
AN - SCOPUS:84968546872
SN - 1059-1524
VL - 27
SP - 1561
EP - 1569
JO - Molecular biology of the cell
JF - Molecular biology of the cell
IS - 10
ER -