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An Anaerobic Biosensor Assay for the Detection of Mercury and Cadmium

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A genetically encoded fluorescent sensor for manganese(II), engineered from lanmodulin.

Jennifer Park¹, Michael B Cleary², Danyang Li³

¹Department of Chemistry, The Pennsylvania State University, University Park, PA 16802.

Proceedings of the National Academy of Sciences of the United States of America

|December 12, 2022

Summary

This summary is machine-generated.

Researchers engineered fluorescent sensors from lanmodulin to detect manganese (II) ions, overcoming challenges in metal-binding site design. These tools enable visualization of manganese in live cells, advancing studies in biology and neurobiology.

Keywords:

Irving-Williams series bioinorganic chemistry fluorescent sensors metal selectivity protein engineering

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Area of Science:

Biochemistry
Chemical Biology
Protein Engineering

Background:

Designing selective metal-binding sites is challenging, especially for manganese (II) due to its low ligand affinity.
A lack of tools hinders the study of manganese physiology in live cells, impacting fields like photosynthesis and neurobiology.

Purpose of the Study:

To develop genetically encoded fluorescent sensors for selective manganese (II) detection.
To re-engineer the lanthanide-binding protein, lanmodulin, for manganese sensing.

Main Methods:

Rational re-engineering of lanmodulin to create MnLaMP1 and MnLaMP2 sensors.
In vitro and in vivo detection of manganese (II) using the developed sensors.
Application of sensors to visualize bacterial labile manganese pools.

Main Results:

Developed two fluorescent sensors, MnLaMP1 and MnLaMP2, with effective dissociation constants (Kd) for Mn(II) of 29 and 7 µM, respectively.
Demonstrated selective detection of Mn(II) in vitro and in vivo, defying the Irving-Williams series.
Visualized the kinetics of labile manganese pools in bacteria.

Conclusions:

Lanmodulin serves as a versatile scaffold for creating selective protein-based biosensors and chelators.
The engineered sensors provide crucial tools for studying manganese physiology in live cellular environments.
Understanding coordinated solvent and hydrophobic interactions is key to sensor selectivity.