biostudies-literatureTan WNational Institute of Allergy and Infectious DiseasesNational Cancer InstituteNCI NIH HHSDivision of Materials Research6709-6713https://www.ebi.ac.uk/biostudies/studies/S-EPMC9069992biostudies-literatureUnknown144(15)The Golgi apparatus (GA) is the hub of intracellular trafficking, but selectively targeting GA remains a challenge. We show an unconventional types of peptide thioesters, consisting of an aminoethyl thioester and acting as substrates of thioesterases, for instantly targeting the GA of cells. The peptide thioesters, above or below their critical micelle concentrations, enter cells mainly via caveolin-mediated endocytosis or macropinocytosis, respectively. After being hydrolyzed by GA-associated thioesterases, the resulting thiopeptides form dimers and accumulate in the GA. After saturating the GA, the thiopeptides are enriched in the endoplasmic reticulum (ER). Their buildup in ER and GA disrupts protein trafficking, thus leading to cell death via multiple pathways. The peptide thioesters target the GA of a wide variety of cells, including human, murine, and <i>Drosophila</i> cells. Changing d-diphenylalanine to l-diphenylalanine in the peptide maintains the GA-targeting ability. In addition, targeting GA redirects protein (e.g., NRAS) distribution. This work illustrates a thioesterase-responsive and redox-active molecular platform for targeting the GA and controlling cell fates.Journal of the American Chemical SocietyEnzyme-Responsive Peptide Thioesters for Targeting Golgi Apparatus.PMC9069992DMR-2011846CA142746AI141386R01 CA142746Luo HRQuinones-Frias MCRodal ATan WZhang YXu BHsu AYHong PZhang QfalseEnzyme-Responsive Peptide Thioesters for Targeting Golgi Apparatus.The Golgi apparatus (GA) is the hub of intracellular trafficking, but selectively targeting GA remains a challenge. We show an unconventional types of peptide thioesters, consisting of an aminoethyl thioester and acting as substrates of thioesterases, for instantly targeting the GA of cells. The peptide thioesters, above or below their critical micelle concentrations, enter cells mainly via caveolin-mediated endocytosis or macropinocytosis, respectively. After being hydrolyzed by GA-associated thioesterases, the resulting thiopeptides form dimers and accumulate in the GA. After saturating the GA, the thiopeptides are enriched in the endoplasmic reticulum (ER). Their buildup in ER and GA disrupts protein trafficking, thus leading to cell death via multiple pathways. The peptide thioesters target the GA of a wide variety of cells, including human, murine, and <i>Drosophila</i> cells. Changing d-diphenylalanine to l-diphenylalanine in the peptide maintains the GA-targeting ability. In addition, targeting GA redirects protein (e.g., NRAS) distribution. This work illustrates a thioesterase-responsive and redox-active molecular platform for targeting the GA and controlling cell fates.2022-01-01T00:00:00Z2022 Apr2025-04-04T11:39:43.622Z2025-04-04T11:39:43.622ZS-EPMC90699923540459910.1021/jacs.2c02238