Recent Progresses of Peptide Fluorescent Probes for Protein Analysis in Living Cells

The accurate and sensitive detection of proteins is crucial for unraveling biological intricacies. Fluorescent probes have emerged as a preferred tool in protein analysis due to their high specificity and nondestructive imaging capabilities. Recent advancements have shown that multifunctional peptide-based fluorescent probes can significantly improve protein analysis performance in living cells. By focusing on the surface properties or chemical characteristics of proteins, we have categorized the design strategies of probes into three types: (1) weak interaction-probe binding predominantly through noncovalent bonds within specialized pockets of targeted protein mainly; (2) strong interaction-probes attach via covalent bond to protein active groups or utilizing protein’s catalytic activity to alter fluorescence signals; (3) combined weak and strong interactions─probes first bind to the targeted proteins through noncovalent interactions and subsequently trigger proximity-induced covalent bond reactions. This review systematically summarizes peptide-based fluorescent probes’ design principles and recognition mechanisms and their applications in living cells. It also discusses the challenges and opportunities in this field, providing insights to promote the innovative development of protein-targeted peptide fluorescent probes.