Tuning the metal binding site specificity of a fluorescent sensor protein: from copper to zinc and back.
We previously reported the development of high affinity Zn(2+) FRET sensors based on the Zn(2+)-mediated interaction between the CXXC motifs present in the copper chaperone proteins ATOX1 and WD4. By systematically substituting several of these cysteines for methionines, we constructed sensor variants that retain a high affinity for Cu(+), while effectively abolishing their ability to form stable tetrahedral Zn(2+) complexes.
Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. m.s.t.koay@utwente.nl
Melissa S
MS
Koay
Brian M G
BM
Janssen
Maarten
M
Merkx
D000595:Amino Acid Sequence; D001665:Binding Sites; D003300:Copper / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D056511:Metallochaperones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering; D013379:Substrate Specificity; D015032:Zinc / Q000378:metabolism*
D005456:Fluorescent Dyes; D056511:Metallochaperones; D003300:Copper; D015032:Zinc