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BODIPY-Patatin assay proteomics

ABSTRACT: The potato protein patatin embeds bioactive peptides that require targeted hydrolysis to be released as promising food additives. This study presents a patatin-specific protease assay for assessing a wide range of protease activities in high-throughput format. Conjugating patatin to the amine reactive fluorogenic BODIPY FL dye provided a stable protease substrate with efficient homo-FRET quenching at a low degree (7-8) of labeling. Compared to commercial BODIPY-casein, BODIPY-patatin provided higher fluorescence enhancement (by de-quenching) at high protease concentrations, while the sensitivity was generally comparable for both highly specific (e.g. Trypsin) and industrial relevant proteases (e.g. Alcalase and Neutrase) at low doses. For Chymotrypsin, BODIPY-patatin provided a 39% response improvement at 5 ng dose. Peptide-centric mass spectrometry analyses identified several BODIPY-labeling sites with varying occupancies in patatin, indicating heterogenous labeling under the applied conjugation conditions. BODIPY-labeled patatin complements commercial BODIPY-labeled casein as a globular, plant-based alternative for screening of proteolytic activity.

INSTRUMENT(S):

ORGANISM(S): Solanum Tuberosum (potato)

TISSUE(S): Plant Cell

SUBMITTER: Simon Gregersen Echers

LAB HEAD: Simon Gregersen Echers

PROVIDER: PXD049376 | Pride | 2025-10-22

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	Alc_small_BODIPY_site10.zip	Other
	Alc_small_BODIPY_site100.zip	Other
	Alcalase_Full.zip	Other
	Alcalase_small.zip	Other
	BODIPY-Pat_Alc_10_SGR_PROVIDE.raw	Raw

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