RRC ID 39509
Author Morikawa TJ, Fujita H, Kitamura A, Horio T, Yamamoto J, Kinjo M, Sasaki A, Machiyama H, Yoshizawa K, Ichimura T, Imada K, Nagai T, Watanabe TM.
Title Dependence of fluorescent protein brightness on protein concentration in solution and enhancement of it.
Journal Sci Rep
Abstract Fluorescent proteins have been widely used in biology because of their compatibility and varied applications in living specimens. Fluorescent proteins are often undesirably sensitive to intracellular conditions such as pH and ion concentration, generating considerable issues at times. However, harnessing these intrinsic sensitivities can help develop functional probes. In this study, we found that the fluorescence of yellow fluorescent protein (YFP) depends on the protein concentration in the solution and that this dependence can be enhanced by adding a glycine residue in to the YFP; we applied this finding to construct an intracellular protein-crowding sensor. A Förster resonance energy transfer (FRET) pair, involving a cyan fluorescent protein (CFP) insensitive to protein concentration and a glycine-inserted YFP, works as a genetically encoded probe to evaluate intracellular crowding. By measuring the fluorescence of the present FRET probe, we were able to detect dynamic changes in protein crowding in living cells.
Volume 6
Pages 22342
Published 2016-3-9
DOI 10.1038/srep22342
PII srep22342
PMID 26956628
PMC PMC4783657
MeSH Fluorescence Resonance Energy Transfer Glycine / genetics Luminescent Measurements / methods* Luminescent Proteins / analysis* Luminescent Proteins / genetics Recombinant Proteins / analysis Recombinant Proteins / genetics Solutions / chemistry*
IF 3.998
Times Cited 23
DNA material pGimRET (RDB14204) pPAL7_GimRET (RDB14203).
Human and Animal Cells NB2a(RCB2639)