State-dependent motion of a genetically encoded fluorescent biosensor

Genetically encoded biosensors can measure biochemical properties such as small-molecule concentrations with single-cell resolution, even in vivo. Despite their utility, these sensors are "black boxes": Very little is known about the structures of their low- and high-fluorescence states or what features are required to transition between them. We used LiLac, a lactate biosensor with a quantitative fluorescence-lifetime readout, as a model system to address these questions. X-ray crystal structures and engineered high-affinity metal bridges demonstrate that LiLac exhibits a large interdomain twist motion that pulls the fluorescent protein away from a "sealed," high-lifetime state in the absence of lactate to a "cracked," low-lifetime state in its presence. Understanding the structures and dynamics of LiLac will help to think about and engineer other fluorescent biosensors.

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Publication Details

Journal
Proceedings of the National Academy of Sciences
Published
2025-03-06
DOI
https://doi.org/10.1073/pnas.2426324122
Citations
7
Primary Topic
Gene Regulatory Network Analysis
Type
article
Field-Weighted Citation Impact
5.64

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article

State-dependent motion of a genetically encoded fluorescent biosensor

Joseph A. Ambarian, Samantha M. Horwitz, Gary Yellen, Erica Kim et al.
7 citations
Proceedings of the National Academy of Sciences
Gene Regulatory Network Analysis
5.64
article

State-dependent motion of a genetically encoded fluorescent biosensor

Joseph A. Ambarian, Samantha M. Horwitz, Gary Yellen, Erica Kim, Paul C. Rosen, Daniel J. Brooks, Katherine M. Davis, Lidia M Waidmann
article en
7 citations

Abstract

Genetically encoded biosensors can measure biochemical properties such as small-molecule concentrations with single-cell resolution, even in vivo. Despite their utility, these sensors are "black boxes": Very little is known about the structures of their low- and high-fluorescence states or what features are required to transition between them. We used LiLac, a lactate biosensor with a quantitative fluorescence-lifetime readout, as a model system to address these questions. X-ray crystal structures and engineered high-affinity metal bridges demonstrate that LiLac exhibits a large interdomain twist motion that pulls the fluorescent protein away from a "sealed," high-lifetime state in the absence of lactate to a "cracked," low-lifetime state in its presence. Understanding the structures and dynamics of LiLac will help to think about and engineer other fluorescent biosensors.

Proceedings of the National Academy of SciencesVol. 122(10)
Harvard University (US), Emory University (US), Massachusetts Institute of Technology (US)
National Science Foundation, Emory University, Department of Neurobiology, Harvard Medical School, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Office of Science, NIH Office of the Director, National Cancer Institute, National Institute of General Medical Sciences, Biological and Environmental Research
Openalex Percentile: Top 4%
Gene Regulatory Network Analysis
5.64
DOIOpen Access PDFOpenAlex
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State-dependent motion of a genetically encoded fluorescent biosensor — Joseph A. Ambarian, Samantha M. Horwitz, et al. · Proceedings of the National Academy of Sciences (2025) | TGRS Research Map | TGRS