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Next Generation Protein Sequencing

ABSTRACT: NGPS is a method for de-novo, full-length protein sequencing in high throughput. The method is based on cleavage of the protein at semi-random sites by microwave-assisted acid hydrolysis (MAAH), enrichment of LC-MS/MS amenable peptides from the hydrolysate by solid-phase-extraction, LC-MS/MS analysis, de-novo long peptide tag sequencing of resulting peptides and assembly of peptide tags into consensus contigs.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Equus Caballus (horse) Bos Taurus (bovine)

SUBMITTER: Alon Savidor

LAB HEAD: Yishai Levin

PROVIDER: PXD003804 | Pride | 2017-04-03

REPOSITORIES: Pride

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Database-independent Protein Sequencing (DiPS) Enables Full-length <i>de Novo</i> Protein and Antibody Sequence Determination.

Savidor Alon A Barzilay Rotem R Elinger Dalia D Yarden Yosef Y Lindzen Moshit M Gabashvili Alexandra A Adiv Tal Ophir O Levin Yishai Y

Molecular & cellular proteomics : MCP 20170327 6

Traditional "bottom-up" proteomic approaches use proteolytic digestion, LC-MS/MS, and database searching to elucidate peptide identities and their parent proteins. Protein sequences absent from the database cannot be identified, and even if present in the database, complete sequence coverage is rarely achieved even for the most abundant proteins in the sample. Thus, sequencing of unknown proteins such as antibodies or constituents of metaproteomes remains a challenging problem. To date, there is ...[more]

PMID: 28348172

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