Dataset Information

One-step Isolation of Protein C-terminal Peptides from Protease-digested Proteins by Metal Oxide-based Ligand Exchange Chromatography

ABSTRACT: We have developed a one-step method to isolate protein C-terminal peptides from V8-digested proteins by metal oxide-based ligand-exchange (MOLEX) chromatography. V8 protease cleaves the C-terminal side of Asp and Glu, and the resulting digested peptide has two carboxy groups at the C-terminus, whereas the protein C-terminal peptide has only one α-carboxy group. In MOLEX chromatography, a stable chelate is formed between dicarboxylates and metal atoms, so that the non-terminal (i.e., internal) peptide is retained, but the protein C-terminal peptide flows through the MOLEX column. By optimizing the conditions of V8 protease digestion and MOLEX chromatographic isolation, a total of 1619 protein C-termini from 30 μg of peptides derived from human HeLa cells could be identified in 10 min of the isolation step. Furthermore, using 200 µg of peptide, we identified 2202 protein C-termini, which was the largest data set with the least amount of sample in protein C-terminomics. These results indicated that this method is unprecedentedly simple and sensitive for protein C-terminal peptide enrichment.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Yasushi Ishihama

PROVIDER: PXD027960 | JPOST Repository | Tue Dec 28 00:00:00 GMT 2021

REPOSITORIES: jPOST

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Dataset's files

Source:

			Action	DRS
	181217HN_26_CeO2_acetate_pH5.wiff.scan	Wiff
	181217HN_27_CeO2_acetate_pH4.wiff.scan	Wiff
	181217HN_28_CeO2_acetate_pH3.wiff.scan	Wiff
	181217HN_29_CeO2_acetate_pH2.wiff.scan	Wiff
	190415HN_10_CeO2_30mMacetate_pH3.wiff.scan	Wiff

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One-Step Isolation of Protein C-Terminal Peptides from V8 Protease-Digested Proteins by Metal Oxide-Based Ligand-Exchange Chromatography.

Nishida Hiroshi H Ishihama Yasushi Y

Analytical chemistry 20211228 2

We have developed a one-step method to isolate protein C-terminal peptides from V8 protease-digested proteins by metal oxide-based ligand-exchange (MOLEX) chromatography. V8 protease cleaves the C-terminal side of Asp and Glu, affording a digested peptide with two carboxy groups at the C-terminus, whereas the protein C-terminal peptide has only one α-carboxy group. In MOLEX chromatography, a stable chelate is formed between dicarboxylates and metal atoms, so that the nonterminal (i.e., internal) ...[more]

PMID: 34962382

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