ArticleApplied and environmental microbiology2007
Design and application of highly responsive fluorescence resonance energy transfer biosensors for detection of sugar in living Saccharomyces cerevisiae cells.
Article in Applied and environmental microbiology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 45 citations in OpenAlex.
- Microfluidic Single-Cell Analytics.Advances in biochemical engineering/biotechnology · 2022Article
- FRET Microscopy in Yeast.Biosensors · 2019Review
- Self Assembled Recombinant Proteins on Metallic Nanoparticles As Bimodal Imaging Probes.JOM (Warrendale, Pa. : 1989) · 2019Article
- A synthetic microbial biosensor for high-throughput screening of lactam biocatalysts.Nature communications · 2018Article
- Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer.Journal of visualized experiments : JoVE · 2016Article
- Glucose recognition proteins for glucose sensing at physiological concentrations and temperatures.ACS chemical biology · 2014Article
- Microbial biosensors: engineered microorganisms as the sensing machinery.Sensors (Basel, Switzerland) · 2013Review
- Generating in vivo cloning vectors for parallel cloning of large gene clusters by homologous recombination.PloS one · 2013Article
- Nucleic acid sandwich hybridization assay with quantum dot-induced fluorescence resonance energy transfer for pathogen detection.Sensors (Basel, Switzerland) · 2012Article
- Mining the Sinorhizobium meliloti transportome to develop FRET biosensors for sugars, dicarboxylates and cyclic polyols.PloS one · 2012Article
- A bacteria colony-based screen for optimal linker combinations in genetically encoded biosensors.BMC biotechnology · 2011Article
- Optical sensors for measuring dynamic changes of cytosolic metabolite levels in yeast.Nature protocols · 2011Article
- Visualizing dynamic interaction between calmodulin and calmodulin-related kinases via a monitoring method in live mammalian cells.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Tolerance of the Ralstonia eutropha class I polyhydroxyalkanoate synthase for translational fusions to its C terminus reveals a new mode of functional display.Applied and environmental microbiology · 2009Article
- Quantitative imaging for discovery and assembly of the metabo-regulome.The New phytologist · 2008Review
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A protein sensor with a highly responsive fluorescence resonance energy transfer (FRET) signal for sensing sugars in living Saccharomyces cerevisiae cells was developed by combinatorial engineering of the domain linker and the binding protein moiety. Although FRET sensors based on microbial binding proteins have previously been created for visualizing various sugars in vivo, such sensors are limited due to a weak signal intensity and a narrow dynamic range. In the present study, the length and composition of the linker moiety of a FRET-based sensor consisting of CFP-linker(1)-maltose-binding protein-linker(2)-YFP were redesigned, which resulted in a 10-fold-higher signal intensity. Molecular modeling of the composite linker moieties, including the connecting peptide and terminal regions of the flanking proteins, suggested that an ordered helical structure was preferable for tighter coupling of the conformational change of the binding proteins to the FRET response. When the binding site residue Trp62 of the maltose-binding protein was diversified by saturation mutagenesis, the Leu mutant exhibited an increased binding constant (82 microM) accompanied by further improvement in the signal intensity. Finally, the maltose sensor with optimized linkers was redesigned to create a sugar sensor with a new specificity and a wide dynamic range. When the optimized maltose sensors were employed as in vivo sensors, highly responsive FRET images were generated from real-time analysis of maltose uptake of Saccharomyces cerevisiae (baker's yeast).
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