title: Genetically encoded FRET sensors to monitor intracellular Zn2+ homeostasis.
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.
authors.affiliation: Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands.
authors.forename: Jan L
authors.initials: JL
authors.lastname: Vinkenborg
authors.forename: Tamara J
authors.initials: TJ
authors.lastname: Nicolson
authors.forename: Elisa A
authors.initials: EA
authors.lastname: Bellomo
authors.forename: Melissa S
authors.initials: MS
authors.lastname: Koay
authors.forename: Guy A
authors.initials: GA
authors.lastname: Rutter
authors.forename: Maarten
authors.initials: M
authors.lastname: Merkx
mesh_terms: 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*
chemical_list: D011994:Recombinant Proteins; D015032:Zinc