Ratiometric biosensors based on dimerization-dependent fluorescent protein exchange

Yidan Ding; Jiao Li; Jhon Ralph Enterina; Yi Shen; Issan Zhang; Paul H Tewson; Gary C H Mo; Jin Zhang; Anne Marie Quinn; Thomas E Hughes; Dusica Maysinger; Spencer C Alford; Yan Zhang; Robert E Campbell

doi:10.1038/nmeth.3261

Ratiometric biosensors based on dimerization-dependent fluorescent protein exchange

Nat Methods. 2015 Mar;12(3):195-8. doi: 10.1038/nmeth.3261. Epub 2015 Jan 26.

Authors

Affiliations

¹ Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada.
² State Key Laboratory of Biomembrane and Membrane Biotechnology, College of Life Sciences, PKU (Peking University)-IDG (International Data Group)/McGovern Institute for Brain Research, Peking University, Beijing, China.
³ Department of Pharmacology and Therapeutics, McGill University, Montreal, Quebec, Canada.
⁴ Montana Molecular, Bozeman, Montana, USA.
⁵ Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
⁶ 1] Montana Molecular, Bozeman, Montana, USA. [2] Department of Cell Biology and Neuroscience, Montana State University, Bozeman, Montana, USA.
⁷ 1] Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada. [2] Department of Bioengineering, Stanford University, Stanford, California, USA.

Abstract

We have developed a versatile new class of genetically encoded fluorescent biosensor based on reversible exchange of the heterodimeric partners of green and red dimerization-dependent fluorescent proteins. We demonstrate the use of this strategy to construct both intermolecular and intramolecular ratiometric biosensors for qualitative imaging of caspase activity, Ca(2+) concentration dynamics and other second-messenger signaling activities.

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

Biosensing Techniques / methods*
Calcium / metabolism
Calmodulin / metabolism
Caspase 3 / genetics
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism*
HeLa Cells
Humans
Luminescent Proteins / genetics
Luminescent Proteins / metabolism*
Molecular Imaging / methods
Protein Multimerization
Red Fluorescent Protein

Substances

Calmodulin
Luminescent Proteins
Green Fluorescent Proteins
Caspase 3
Calcium

Abstract

Publication types

MeSH terms

Substances

Grants and funding