Reply to Goodman et al.: Imaging protein synthesis with puromycin and the subcellular localization of puromycin–polypeptide conjugates

Abstract

The antibiotic puromycin attaches covalently to the C terminus of nascent polypeptide chains, causing their premature termination and rapid dissociation from ribosomes. This mechanism allows the use of puromycin to assay protein synthesis in cells by detection of polypeptide–puromycin conjugates with tritiated puromycin (1), antipuromycin antibodies (2), or clickable O-propargyl-puromycin (3). Although antipuromycin antibodies work well on blots and for FACS analysis (2), they reveal a subcellular pattern of puromycin conjugates that is significantly different from the pattern of the methionine analog homopropargylglycine (4), O-propargyl-puromycin (3), and tritiated puromycin (1) (see below). The letter by … [↵][1]1E-mail: asalic{at}hms.harvard.edu. [1]: #xref-corresp-1-1