Abstract：
:Biomolecular dynamics and stability are predominantly investigated in vitro and extrapolated to explain function in the living cell. We present fast relaxation imaging (FreI), which combines fluorescence microscopy and temperature jumps to probe biomolecular dynamics and stability inside a single living cell with high spatiotemporal resolution. We demonstrated the method by measuring the reversible fast folding kinetics as well as folding thermodynamics of a fluorescence resonance energy transfer (FRET) probe-labeled phosphoglycerate kinase construct in two human cell lines. Comparison with in vitro experiments at 23-49 degrees C showed that the cell environment influences protein stability and folding rate. FReI should also be applicable to the study of protein-protein interactions and heat-shock responses as well as to comparative studies of cell populations or whole organisms.
journal_name
Nat Methodsjournal_title
Nature methodsauthors
Ebbinghaus S,Dhar A,McDonald JD,Gruebele Mdoi
10.1038/nmeth.1435subject
Has Abstractpub_date
2010-04-01 00:00:00pages
319-23issue
4eissn
1548-7091issn
1548-7105pii
nmeth.1435journal_volume
7pub_type
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