Proteomics

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Reversible Click Chemistry Tag for Universal Proteome Sample Preparation

ABSTRACT: Here, we introduce a reversible protein tag, ProMTag, that enables whole proteome capture, cleanup, and release of intact proteins for top-down analysis. Alternatively, the addition of a novel Trypsin derivative to the workflow generates peptides for bottom-up analysis. We show that the ProMTag workflow yields >90% for intact proteins and >85% for proteome digests. For top-down analysis, ProMTag cleanup improves resolution on 2D gels; for bottom-up exploration, this methodology produced reproducible mass spectrometry results, demonstrating that the ProMTag method is a truly universal approach that produces high quality proteome samples compatible with multiple downstream analytical techniques.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Gallus Gallus (chicken) Burkholderia Sp. 2332 Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Jonathan Minden  

LAB HEAD: Amber Lucas

PROVIDER: PXD027799 | Pride | 2022-02-14

REPOSITORIES: pride

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Publications

Reversible Click Chemistry Tag for Universal Proteome Sample Preparation for Top-Down and Bottom-Up Analysis.

Biedka Stephanie S   Schmidt Brigitte F BF   Frey Nolan M NM   Boothman Sarah M SM   Minden Jonathan S JS   Wilson Amber Lee AL  

Journal of proteome research 20210915 10

Successful proteome analysis requires reliable sample preparation beginning with protein solubilization and ending with a sample free of contaminants, ready for downstream analysis. Most proteome sample preparation technologies utilize precipitation or filter-based separation, both of which have significant disadvantages. None of the current technologies are able to prepare both intact proteins or digested peptides. Here, we introduce a reversible protein tag, ProMTag, that enables whole proteom  ...[more]

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