Abstract
We developed genetically encoded fluorescence resonance energy transfer (FRET)-based sensors that display a large ratiometric change upon Zn(2+) binding, have affinities that span the pico- to nanomolar range and can readily be targeted to subcellular organelles. Using this sensor toolbox we found that cytosolic Zn(2+) was buffered at 0.4 nM in pancreatic beta cells, and we found substantially higher Zn(2+) concentrations in insulin-containing secretory vesicles.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Animals
-
Biological Assay / methods*
-
Cell Line
-
Fluorescence Resonance Energy Transfer / methods*
-
Homeostasis / physiology*
-
Insulin-Secreting Cells / metabolism
-
Molecular Probe Techniques*
-
Protein Engineering / methods*
-
Rats
-
Recombinant Proteins / analysis
-
Recombinant Proteins / metabolism*
-
Zinc / analysis
-
Zinc / metabolism*
Substances
-
Recombinant Proteins
-
Zinc