Illuminating biological processes through site-specific protein labeling

• Published in: Chemical Society reviews Vol. 44; no. 11; p. 3405
• Main Authors: Zhang, Gong, Zheng, Siqi, Liu, Haiping, Chen, Peng R
• Format: Journal Article
• Language: English
• Published: England 07-06-2015
• Subjects: Humans; Models, Biological; Models, Molecular; Molecular Probe Techniques; Proteins - analysis; Proteins - chemistry; Proteins - metabolism; Proteins - physiology
• ISSN: 1460-4744
• DOI: 10.1039/c4cs00393d

Coupling genetically encoded peptide tags or unnatural amino acids (UAAs) with bioorthogonal reactions allows for precise control over the protein-labeling sites as well as the wide choice of labeling dyes. However, the value of these site-specific protein labeling strategies in a real biology setting, particularly their advantages over conventional labeling methods including fluorescent proteins (FPs), remains to be fully demonstrated. In this tutorial review, we first introduce various strategies for site-specific protein labeling that utilize artificial peptide sequences or genetically encoded UAAs as the labeling handle. Emphasis will be placed on introducing the advantages of protein site-specific labeling techniques as well as their applications in solving biological problems, particularly as to why a site-specific protein labeling approach is needed. Finally, beyond the widely used single site-specific labeling methods, the recently emerged dual site-specific protein labeling strategies will be introduced together with their fast-growing potential in illustrating biological processes.