| Engineering Ca2+/calmodulin-mediated modulation of protein translocation by overlapping binding and signaling peptide sequences. | |
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MedLine Citation:
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PMID: 20167369 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Protein translocation is used by cells to regulate protein activity in time and space. Synthetic systems have studied the effect of second messengers and exogenous chemicals on translocation, and have used translocation-based sensors to monitor unrelated pathways such as caspase activity. We have created a synthetic Ca2+-inducible protein using calmodulin binding peptides that selectively reveal nuclear localization and export signals in low Ca2+ (0 microM) and high Ca2+ (10 microM) environments, respectively. Experiments in live cells showed that our construct translocates between the nucleolus and plasma membrane with time constants of approximately 2 h. Further, a single amino acid mutation (Cys20Ala) in our construct prevented translocation to the plasma membrane and instead targeted it the mitochondria as predicted by bioinformatic analysis. Lastly, we studied the effect of cell line, Ca2+ concentration, chemical inhibitors, and cell morphology on translocation and found these conditions affected the rate, extent and direction of translocation. Our work demonstrates the feasibility of engineering Ca2+/calmodulin-mediated modulation of protein translocation and suggests that more natural analogs may exist. |
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Authors:
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Evan Mills; Kevin Truong |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-02-18 |
Journal Detail:
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Title: Cell calcium Volume: 47 ISSN: 1532-1991 ISO Abbreviation: Cell Calcium Publication Date: 2010 Apr |
Date Detail:
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Created Date: 2010-04-05 Completed Date: 2010-09-07 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8006226 Medline TA: Cell Calcium Country: Netherlands |
Other Details:
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Languages: eng Pagination: 369-77 Citation Subset: IM |
Copyright Information:
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2010 Elsevier Ltd. All rights reserved. |
Affiliation:
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Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario M5S3G9, Canada. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Active Transport, Cell Nucleus Animals COS Cells Calcium Signaling* Calmodulin / metabolism Cell Membrane / metabolism Cell Nucleus / metabolism* Cercopithecus aethiops Computational Biology Mitochondria / metabolism Mutagenesis, Site-Directed Mutation / genetics Nuclear Localization Signals / genetics Nucleolus Organizer Region / metabolism Peptide Fragments / genetics, metabolism* Protein Binding Protein Engineering Recombinant Proteins / genetics, metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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81262//Canadian Institutes of Health Research |
| Chemical | |
Reg. No./Substance:
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0/Calmodulin; 0/Nuclear Localization Signals; 0/Peptide Fragments; 0/Recombinant Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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