Proteomics

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Profiling of Native and Protease_Generated Protein Amino Termini from Formalin-Fixed, Paraffin-Embedded Tissues


ABSTRACT: Dysregulated proteolysis represents a hallmark of numerous diseases. In recent years, increasing number of studies has begun looking at the protein termini in hope to unveil the physiological and pathological functions of proteases in clinical research. However, the availability of cryopreserved tissue specimens is often limited. Alternatively, formalin-fixed, paraffin-embedded (FFPE) tissues offer an invaluable resource for clinical research. Pathologically relevant tissues are often stored as FFPE, which represent the most abundant resource of archived human specimens. In this study, we established a robust workflow to investigate native and protease-generated protein N-termini from FFPE specimens. We demonstrate a comparable N-terminomes of cryopreserved and formalin fixed tissue, thereby showing that formalin fixation / paraffin embedment does not proteolytically damage proteins. Accordingly, FFPE specimens are fully amenable to N-terminal analysis. Moreover, we demonstrate feasibility of FFPE-degradomics in a quantitative N-terminomic study of FFPE liver specimens from cathepsin L deficient or wild-type mice. Using a machine learning approach in combination with the previously determined cathepsin L specificity, we successfully identified a number of potential cathepsin L cleavage sites. Our study establishes FFPE specimens as a valuable alternative to cryopreserved tissues for degradomic studies.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Liver

SUBMITTER: Zon Lai  

LAB HEAD: Oliver Schilling

PROVIDER: PXD002847 | Pride | 2016-04-25

REPOSITORIES: Pride

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Publications

Formalin-Fixed, Paraffin-Embedded Tissues (FFPE) as a Robust Source for the Profiling of Native and Protease-Generated Protein Amino Termini.

Lai Zon Weng ZW   Weisser Juliane J   Nilse Lars L   Costa Fabrizio F   Keller Eva E   Tholen Martina M   Kizhakkedathu Jayachandran N JN   Biniossek Martin M   Bronsert Peter P   Schilling Oliver O  

Molecular & cellular proteomics : MCP 20160417 6


Dysregulated proteolysis represents a hallmark of numerous diseases. In recent years, increasing number of studies has begun looking at the protein termini in hope to unveil the physiological and pathological functions of proteases in clinical research. However, the availability of cryopreserved tissue specimens is often limited. Alternatively, formalin-fixed, paraffin-embedded (FFPE) tissues offer an invaluable resource for clinical research. Pathologically relevant tissues are often stored as  ...[more]

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