A changed cameleon | Nature Reviews Molecular Cell Biology

Access through your institution Buy or subscribe Amy Palmer in the Tsien laboratory started with the original cameleon construct — two fluorescent proteins (cyan fluorescent protein (CFP)

and citrine) separated by calmodulin (CaM) and a CaM-binding peptide. In the presence of Ca2+, CaM interacts with the CaM-binding peptide, and CFP emission decreases as citrine emission

increases, which is indicative of increased fluorescence resonance energy transfer (FRET). Their aim was to change this cameleon to decrease its perturbation by endogenous CaM and to vary

its Ca2+ affinity. Using previously obtained structural data, Palmer targeted salt-bridge interactions between CaM and the CaM-binding peptide. Compared to the wild-type peptide, a mutant

peptide with four charge reversals had a 10,000-fold lower affinity for wild-type CaM. Palmer then made compensatory charge reversals in CaM with the aim of restoring its affinity for the

mutant peptide. She named the mutant CaM and peptide pair Design 1 (D1), and cloned it between CFP and citrine to produce an altered cameleon. This is a preview of subscription content,

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OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support REFERENCES ORIGINAL RESEARCH PAPER * Palmer, A. E. et al. Bcl-2-mediated alterations in

endoplasmic reticulum Ca2+ analyzed with an improved genetically encoded fluorescent sensor. _Proc. Natl Acad. Sci. USA_ 101, 17404–17409 (2004) Article  CAS  Google Scholar  FURTHER

READING * Rudolf, R. et al. Looking forward to seeing calcium. _Nature Rev. Mol. Cell Biol._ 4, 579–586 (2003) Article  CAS  Google Scholar  Download references Authors * Rachel Smallridge

View author publications You can also search for this author inPubMed Google Scholar RELATED LINKS RELATED LINKS WEB SITES John Reed's laboratory Roger Tsien's laboratory RIGHTS

AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Smallridge, R. A changed cameleon. _Nat Rev Mol Cell Biol_ 6, 92 (2005). https://doi.org/10.1038/nrm1584

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