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This protocol from Wang et al. describes a pulse–chase method to investigate autophagic protein degradation through click labeling of long-lived proteins. This is a safer alternative to similar classic methods that use radioactive labeling.
Site-specific labeling of proteins with small fluorophores is advantageous for imaging. Lemke et al. describe how to site-specifically label membrane proteins with organic fluorophores by incorporating non-canonical amino acids via Amber suppression technology.
The combination of a genetically encoded aldehyde tag and optimized labeling method allows high-efficiency, site-specific labeling of tagged proteins after purification or in cell extracts. The authors use the high labeling efficiency for single-molecule measurements of the dynamic interactions between two DNA polymerases and polymerase processivity factor bound to DNA.