Real-time measurement of quorum-sensing signal autoinducer 3OC6HSL by a FRET-based nanosensor.
Quorum sensing (QS) is involved in many important biological functions such as luminescence, antibiotic production, and biofilm formation. The autoinducer N-(3-oxo-hexanoyl)-L-homoserine lactone (3OC6HSL) plays a significant role in the QS system of the marine bacterium Vibrio fischeri. Tracing 3OC6HSL would be significant in studies related to QS signal transduction. Traditional detection of QS signaling molecules has relied on bacterial reporter strains and high-performance liquid chromatography, which are time consuming and have low sensitivity. Because 3OC6HSL binding to LuxR from V. fischeri causes a conformational change, we developed a genetically encoded biosensor based on Förster resonance energy transfer (FRET) by inserting LuxR between the FRET pair YFP/CFP. The detection limit of the sensor was 100 μM. We attained an optimized sensor with 70 % Δratio increase by screening different hydrophobic linkers, and demonstrated the feasibility of this sensor for visualizing 3OC6HSL both in vitro and in vivo.
Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Meilong Road 130, Shanghai, 200237, China.
Chang
C
Zhang
Bang-Ce
BC
Ye
D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000737:chemistry; D048908:Aliivibrio fischeri / Q000254:growth & development*; D001426:Bacterial Proteins / Q000032:analysis*; D015374:Biosensing Techniques / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer; D010675:Pheromones / Q000737:chemistry; D053038:Quorum Sensing; D012097:Repressor Proteins / Q000032:analysis* / Q000378:metabolism; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D015534:Trans-Activators / Q000032:analysis* / Q000378:metabolism
D001426:Bacterial Proteins; D010675:Pheromones; D012097:Repressor Proteins; D015534:Trans-Activators; C064778:LuxR autoinducer binding proteins; C030654:N-(3-oxohexanoyl)-3-aminodihydro-2(3H)-furanone; D015107:4-Butyrolactone