Ca2+ Indicators Based on Computationally Redesigned Calmodulin-Peptide Pairs

Chem Biol. 2006 May;13(5):521-30. doi: 10.1016/j.chembiol.2006.03.007.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Calcium / chemistry
  • Calcium / metabolism*
  • Calmodulin / chemistry
  • Calmodulin / metabolism*
  • Cell Membrane / metabolism
  • Cytosol / metabolism
  • HeLa Cells
  • Humans
  • Models, Molecular
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / metabolism*
  • Protein Binding
  • Subcellular Fractions / metabolism

Substances

  • Calmodulin
  • Peptides
  • Calcium

Associated data

  • GENBANK/DQ479425
  • GENBANK/DQ479426
  • GENBANK/DQ479427
  • GENBANK/DQ479428
  • GENBANK/DQ479429