Ca2+ indicators based on computationally redesigned calmodulin-peptide pairs.
Chem Biol
; 13(5): 521-30, 2006 May.
Article
em En
| MEDLINE
| ID: mdl-16720273
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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Peptídeos
/
Calmodulina
/
Cálcio
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Chem Biol
Ano de publicação:
2006
Tipo de documento:
Article