Proteomics

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LysargiNase Enhances the Protein Identification on FFPE Samples


ABSTRACT: Formalin-Fixed Paraffin-Embedded (FFPE) samples are treasure for proteomic studies of disease. However, their usage is hampered for proteomics study because of the crosslink among proteins, and protein vs nucleic acid. Even worse, other covalent chemical modifications like methylation introduced by formaldehyde interfere trypsin digestion. We applied LysargiNase for the first time in FFPE samples, and systematically compared the samples digested by LysargiNase with commonly used tryptic samples. A total of 33095 peptides and 3559 proteins were identified with LysargiNase and trypsin combined in two replicates. LysargiNase increased peptide identification by 19.3% and protein identification by 13.3% on the basis of trypsin. Consistently, LysargiNase increased C terminal peptide identification by 47.7%. Moreover, LysargiNase showed less missed-cleavage rate (54.5%) at methylated sites than trypsin (74.5%), contributing to the identification of methylated peptides in FFPE samples.

ORGANISM(S): Mus Musculus

SUBMITTER: Ping Xu  

PROVIDER: PXD012738 | iProX | Fri Feb 15 00:00:00 GMT 2019

REPOSITORIES: iProX

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Publications

LysargiNase enhances protein identification on the basis of trypsin on formalin-fixed paraffin-embedded samples.

Liu Shu S   Xu Feng F   Yin Yanan Y   Zhang Junling J   Wang Fuqiang F   Li Yanchang Y   Xu Ping P  

Rapid communications in mass spectrometry : RCM 20190901 17


<h4>Rationale</h4>Formalin-Fixed Paraffin-Embedded (FFPE) samples are valuable for proteomic studies of disease. However, the crosslink among proteins, protein vs nucleic acid, and other covalent chemical modifications like methylation introduced by formaldehyde can interfere with trypsin digestion in proteomics studies. LysargiNase was reported to have a better full-cleavage rate at methylation and b ion coverage than trypsin. The contribution of LysargiNase in the proteomic study of FFPE sampl  ...[more]