Genetically Encoded Near-Infrared Photocatalysis for Proximity Labeling of Subcellular Proteome

Area of Science:

• Cell Biology
• Biochemistry
• Molecular Biology

Background:

• Protein spatial organization is crucial for eukaryotic cellular functions.
• Existing proximity labeling methods use blue light, limiting tissue penetration and causing high background.
• Genetically encoded methods are needed for spatially resolved, proteome-wide protein mapping.

Purpose of the Study:

• To develop a novel near-infrared (NIR) light-activated proximity labeling method.
• To overcome the limitations of blue light activation and antibody-dependent targeting.
• To enable deep tissue protein labeling for live animal applications.

Main Methods:

• Developed FLAPP, a genetically encoded method using engineered fluorogen-activating protein dL5**.
• Utilized near-infrared light activation of a fluorogen-activating protein-fluorogen complex.
• Generated singlet oxygen for in situ labeling of nearby histidine residues.

Main Results:

• FLAPP demonstrates high spatial specificity (96%) in mitochondria and nucleus.
• The method enables efficient protein labeling with deep tissue penetration.
• FLAPP is a genetically encoded, NIR light-activated proximity labeling technology.

Conclusions:

• FLAPP overcomes limitations of blue light and antibody-dependent methods.
• The technique offers precise, spatially resolved protein mapping in deep tissues.
• FLAPP holds significant potential for live animal studies and advanced cellular research.