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Updated: May 15, 2026
Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass
Published on: August 10, 2017
Fluorescence-On Imaging of Golgiphagy with a Golgi Apparatus-Specific Metabolic Lipid Labeling Probe Fluorogenic to
Jipeng Yao1, Shixiong Wen2, Jiahuai Han2,3
1The Key Laboratory for Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, The MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
Abstract:
The Golgi apparatus (GA) plays crucial roles in myriad biological processes, and aberrant Golgiphagy has been implicated in a number of pathological settings. Given the lack of suitable imaging tools for Golgiphagy and the propensity of chemical probes to dissipate from stressed organelles, we herein report Golgi-proRed, a pH probe covalently anchored on the GA inner membrane, for imaging of Golgiphagy by red fluorescence triggered upon autophagic delivery of GA into acidic lysosomes. The probe contains a GA-homing entity of blue-to-green fluorescence, a rhodamine-lactam entity that becomes red-emissive in acidic lysosomes, and a moiety of dibenzocyclooctyne (DBCO) for bioorthogonal conjugation with azido-phospholipids metabolically incorporated into the membrane of GA. Upon Golgiphagy, the membrane-anchored probe is codelivered with GA fragments into lysosomes, yielding red fluorescence that serves as the readout of Golgiphagy. The imaging fidelity for Golgiphagy is confirmed by starvation-induced red fluorescence, lysosome-dependent red fluorescence, and lack of such a signal in ATG5-knockout cells deficient in autophagy. With this method, we observed that obvious Golgiphagy occurred in iBMDM cells treated with nigericin and lipopolysaccharides (LPSs) or infected by Listeria monocytogenes. These results show the use of Golgi-proRed for sensitive detection of Golgiphagy in biological events and to screen Golgiphagy-inducing agents.
