Title |
Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells*
|
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Published in |
Molecular and Cellular Proteomics, September 2018
|
DOI | 10.1074/mcp.tir118.000914 |
Pubmed ID | |
Authors |
Bettina-Maria Keller, Julia Maier, Kathy-Ann Secker, Stefanie-Maria Egetemaier, Yana Parfyonova, Ulrich Rothbauer, Bjoern Traenkle |
Abstract |
Understanding cellular processes requires the determination of dynamic changes in the concentration of genetically non-modified, endogenous proteins, which, to date, is commonly accomplished by endpoint assays in vitro. Molecular probes such as fluorescently labeled nanobodies (chromobodies, CBs) are powerful tools to visualize the dynamic subcellular localization of endogenous proteins in living cells. Here, we employed the dependency of intracellular levels of chromobodies on the amount of their endogenous antigens, a phenomenon, which we termed antigen-mediated CB stabilization (AMCBS), for simultaneous monitoring of time-resolved changes in the concentration and localization of native proteins. To improve the dynamic range of AMCBS we generated turnover-accelerated CBs and demonstrated their application in visualization and quantification of fast reversible changes in antigen concentration upon compound treatment by quantitative live-cell imaging. We expect that this broadly applicable strategy will enable unprecedented insights into the dynamic regulation of proteins, e.g. during cellular signaling, cell differentiation, or upon drug action. |
X Demographics
Geographical breakdown
Country | Count | As % |
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Germany | 2 | 29% |
France | 1 | 14% |
United States | 1 | 14% |
Unknown | 3 | 43% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 4 | 57% |
Scientists | 2 | 29% |
Science communicators (journalists, bloggers, editors) | 1 | 14% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 52 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 16 | 31% |
Researcher | 8 | 15% |
Student > Master | 6 | 12% |
Student > Bachelor | 5 | 10% |
Student > Postgraduate | 3 | 6% |
Other | 2 | 4% |
Unknown | 12 | 23% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 19 | 37% |
Agricultural and Biological Sciences | 5 | 10% |
Chemistry | 3 | 6% |
Immunology and Microbiology | 3 | 6% |
Veterinary Science and Veterinary Medicine | 2 | 4% |
Other | 5 | 10% |
Unknown | 15 | 29% |