Genetically encoded FRET sensors to monitor intracellular Zn2+ homeostasis.
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.
Vinkenborg
Jan L
JL
Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands.
Nicolson
Tamara J
TJ
Bellomo
Elisa A
EA
Koay
Melissa S
MS
Rutter
Guy A
GA
Merkx
Maarten
M
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002460:Cell Line; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006706:Homeostasis / Q000502:physiology*; D050417:Insulin-Secreting Cells / Q000378:metabolism; D015336:Molecular Probe Techniques; D015202:Protein Engineering / Q000379:methods*; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism*; D015032:Zinc / Q000032:analysis / Q000378:metabolism*
D011994:Recombinant Proteins; D015032:Zinc