Green fluorescent protein: structure, folding and chromophore maturation

Area of Science:

• Biochemistry
• Molecular Biology
• Biophysics

Background:

• Green fluorescent protein (GFP) and its variants are crucial tools in cellular biology.
• GFP chromophore formation is a post-translational process requiring only molecular oxygen.
• The maturation involves folding, cyclization, oxidation, and dehydration of the polypeptide backbone.

Purpose of the Study:

• To review proposed mechanisms for GFP chromophore formation.
• To analyze the role of the native state in catalyzing maturation.
• To present the functions of key conserved residues in the maturation process.

Main Methods:

• Detailed examination of experimental studies on GFP maturation.
• Analysis of computational studies investigating chromophore formation mechanisms.
• Discussion of the folding, cyclization, oxidation, and dehydration steps.

Main Results:

• The native state of the polypeptide backbone catalyzes initial cyclization and subsequent chemistry.
• Specific conserved residues (Y66, G67, R96, E222) play critical roles in maturation.
• Proposed mechanisms for chromophore formation are detailed, integrating experimental and computational findings.

Conclusions:

• Understanding GFP maturation mechanisms is essential for advancing fluorescent probe technology.
• Insights gained will facilitate the development of brighter, faster-maturing, and multi-colored fluorescent probes.
• Enhanced fluorescent probes will increase utility across the biomedical sciences.