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Sequential Digestion with Different Proteases

ABSTRACT: We examined three combinations of different proteases for digestion of proteins from TurboID-STING-expressing cells: trypsin alone, Lys-C followed by trypsin, and Glu-C followed by trypsin. First, we tested whether PTS buffer used to solubilize precipitated proteins is compatible with digestion with these proteases. Next, biotinylated peptides were enriched after digestion with the three combinations of different proteases.

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Hidetaka Kosako

PROVIDER: PXD032061 | JPOST Repository | Sat Mar 04 00:00:00 GMT 2023

REPOSITORIES: jPOST

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			Action	DRS
	220225_Tamavidin_OptimizedWF_5_Protease_GluC_Trypsin_FusionIT.pdResult	Other
	220225_Tamavidin_OptimizedWF_5_Protease_GluC_Trypsin_FusionIT_PeptideGroups.txt	Txt
	220225_Tamavidin_OptimizedWF_5_Protease_LysC_Trypsin_FusionIT.pdResult	Other
	220225_Tamavidin_OptimizedWF_5_Protease_LysC_Trypsin_FusionIT_PeptideGroups.txt	Txt
	220225_Tamavidin_OptimizedWF_5_Protease_Trypsin_FusionIT.pdResult	Other

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Sequential Digestion with Different Proteases

Project description:We examined three combinations of different proteases for digestion of proteins from TurboID-STING-expressing cells: trypsin alone, Lys-C followed by trypsin, and Glu-C followed by trypsin. First, we tested whether PTS buffer used to solubilize precipitated proteins is compatible with digestion with these proteases. Next, biotinylated peptides were enriched after digestion with the three combinations of different proteases. We assessed reproducibility of each combination using three technical replicates.

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