Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Ca2+ indicators based on computationally redesigned calmodulin-peptide pairs.

Amy E Palmer1, Marta Giacomello, Tanja Kortemme

  • 1Department of Pharmacology and Howard Hughes Medical Institute, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.

Chemistry & Biology
|May 25, 2006
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Enhancement of RNA Imaging Platforms by the Use of Peptide Nucleic Acid-Based Linkers.

Bio-protocol·2026
Same author

Metal homeostasis is remodeled in response to different quiescence triggers.

bioRxiv : the preprint server for biology·2026
Same author

Zn<sup>2+</sup> transients and signaling in mammalian systems.

Trends in biochemical sciences·2025
Same author

Systematic characterization of zinc in a series of breast cancer cell lines reveals significant changes in zinc homeostasis.

The Journal of biological chemistry·2025
Same author

Structural basis for ring-opening fluorescence by the RhoBAST RNA aptamer.

Nucleic acids research·2025
Same author

Systematic characterization of zinc in a series of breast cancer cell lines reveals significant changes in zinc homeostasis.

bioRxiv : the preprint server for biology·2025
Same journal

The Hedgehog Pathway Effector Smoothened Exhibits Signaling Competency in the Absence of Ciliary Accumulation.

Chemistry & biology·2017
Same journal

DIVERSE System: De Novo Creation of Peptide Tags for Non-enzymatic Covalent Labeling by In Vitro Evolution for Protein Imaging Inside Living Cells.

Chemistry & biology·2015
Same journal

Differential Regulation of Specific Sphingolipids in Colon Cancer Cells during Staurosporine-Induced Apoptosis.

Chemistry & biology·2015
Same journal

Synthetic Peptides as cGMP-Independent Activators of cGMP-Dependent Protein Kinase Iα.

Chemistry & biology·2015
Same journal

Unraveling the B. pseudomallei Heptokinase WcbL: From Structure to Drug Discovery.

Chemistry & biology·2015
Same journal

Vitamin C as Cancer Destroyer, Investigating Sulfhydration, and the Variability in CFTR Interactome.

Chemistry & biology·2015
See all related articles

Researchers engineered new calcium indicators (cameleons) by redesigning protein interfaces. These improved cameleons offer enhanced sensitivity and specificity for sensing calcium ions (Ca2+) across various cellular locations.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • Calmodulin (CaM) is a crucial calcium-binding protein involved in cellular signaling.
  • Genetically encoded calcium indicators (cameleons) are vital tools for monitoring intracellular Ca2+ dynamics.
  • Previous cameleon generations faced limitations with native CaM interference and sensitivity ranges.

Purpose of the Study:

  • To reengineer the calmodulin-binding interface for improved Ca2+ sensing.
  • To develop a new generation of cameleons with enhanced specificity and sensitivity.
  • To broaden the dynamic range and applicability of cameleons in diverse subcellular compartments.

Main Methods:

  • Computational design of complementary "bumps and holes" at the calmodulin-binding interface.

Related Experiment Videos

  • Creation of mutant calmodulin and calmodulin-binding peptides.
  • Generation of genetically encoded calcium indicators (cameleons) incorporating redesigned proteins.
  • Testing Ca2+ sensitivity and dynamic range, including comparisons with native calmodulin.

    • Main Results:

    • Successfully reengineered calmodulin-binding interfaces, creating sensitive and specific mutant protein pairs.
    • Developed second-generation cameleons insensitive to excess native calmodulin.
    • Achieved a 100-fold tunable Ca2+ sensitivity range (0.6–160 µM).
    • Incorporating circularly permuted Venus increased the dynamic range 3- to 5-fold.

    Conclusions:

    • Redesigned cameleons represent a significant advancement in Ca2+ indicator technology.
    • These improved indicators offer greater precision for monitoring Ca2+ in cytoplasm and specific organelles.
    • The enhanced cameleons facilitate more accurate cellular Ca2+ signaling studies in various biological contexts.