Ca2+ indicators based on computationally redesigned calmodulin-peptide pairs

TitleCa2+ indicators based on computationally redesigned calmodulin-peptide pairs
Publication TypeJournal Article
Year of Publication2006
AuthorsPalmer, A. E., Giacomello M., Kortemme T., Hires A. S., Lev-Ram V., Baker D., & Tsien R. Y.
JournalChemistry & biology
Volume13
Issue5
Pagination521-30
Date Published2006 May
ISSN1074-5521
KeywordsAmino Acid Sequence, Calcium, Calmodulin, Cell Membrane, Collaborative Publication, Cytosol, Hela Cells, Humans, Models, Molecular, Molecular Sequence Data, Peptides, Protein Binding, Subcellular Fractions
Abstract

The binding interface of calmodulin and a calmodulin binding peptide were reengineered by computationally designing complementary bumps and holes. This redesign led to the development of sensitive and specific pairs of mutant proteins used to sense Ca(2+) in a second generation of genetically encoded Ca(2+) indicators (cameleons). These cameleons are no longer perturbed by large excesses of native calmodulin, and they display Ca(2+) sensitivities tuned over a 100-fold range (0.6-160 microM). Incorporation of circularly permuted Venus in place of Citrine results in a 3- to 5-fold increase in the dynamic range. These redesigned cameleons show significant improvements over previous versions in the ability to monitor Ca(2+) in the cytoplasm as well as distinct subcellular localizations, such as the plasma membrane of neurons and the mitochondria.

Alternate JournalChem. Biol.
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