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Simple, scalable and sensitive tip-based identification of protease substrates

ABSTRACT: Large-scale N-terminomics using COFRADIC and TAILS has revolutionized protease research, but is disseminated only in few labs. We present an alternative and simple protocol for N-terminal peptide enrichment, based on charge-based fractional diagonal chromatography (ChaFRADIC) and requiring only well-established protein chemistry and a pipette tip. We achieve unprecedented sensitivity quantifying >2000 unique N-terminal peptides from 10 µg per sample, allowing the identification of proteolytic targets and consensus motifs.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Laxmikanth Kollipara

LAB HEAD: Dr. René Peiman Zahedi

PROVIDER: PXD005954 | Pride | 2018-01-24

REPOSITORIES: Pride

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	QExactiveHF02_04436.raw	Raw
	QExactiveHF02_04437.raw	Raw
	QExactiveHF02_04438.raw	Raw
	QExactiveHF02_04439.raw	Raw
	QExactiveHF02_04440.raw	Raw

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Simple, scalable, and ultrasensitive tip-based identification of protease substrates.

Shema Gerta G Nguyen Minh T N MTN Solari Fiorella A FA Loroch Stefan S Venne A Saskia AS Kollipara Laxmikanth L Sickmann Albert A Verhelst Steven H L SHL Zahedi René P RP

Molecular & cellular proteomics : MCP 20180122 4

Proteases are in the center of many diseases, and consequently, proteases and their substrates are important drug targets as represented by an estimated 5-10% of all drugs under development. Mass spectrometry has been an indispensable tool for the discovery of novel protease substrates, particularly through the proteome-scale enrichment of so-called N-terminal peptides representing endogenous protein N termini. Methods such as combined fractional diagonal chromatography (COFRADIC)<sup>1</sup> an ...[more]

PMID: 29358340

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