Variation of Linker Length in Ratiometric Fluorescent Sensor Proteins Allows Rational Tuning of Zn(II) Affinity in the Picomolar to Femtomolar Range

• Published in: Journal of the American Chemical Society Vol. 129; no. 12; pp. 3494 - 3495
• Main Authors: van Dongen, Elisabeth M. W. M, Evers, Toon H, Dekkers, Linda M, Meijer, E. W, Klomp, Leo W. J, Merkx, Maarten
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 28-03-2007
• Subjects: Cross-Linking Reagents - chemistry; Fluorescence Resonance Energy Transfer; Models, Molecular; Peptides - chemistry; Protein Structure, Tertiary; Proteins - chemistry; Zinc - chemistry
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja069105d

Ratiometric fluorescent sensor proteins with a very high and tunable affinity for Zn(II) were created by connecting two fluorescently labeled metal binding domains, CFP−Atox1 and WD4−YFP, using a series of flexible peptide linkers. A simple random-coil model describing the conformational distribution of the linker allowed a quantitative understanding of the effect of the linker length on both the change in emission ratio and the Zn(II) affinity.