Automation of C-terminal sequence analysis of 2D-PAGE separated proteins

Automation of C-terminal sequence analysis of 2D-PAGE separated proteins

Experimental assignment of the protein termini remains essential to define the functional protein structure. Here, we report on the improvement of a proteomic C-terminal sequence analysis method. The approach aims to discriminate the C-terminal peptide in a CNBr-digest where Met-Xxx peptide bonds ar...

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Bibliographic Details
Main Authors: P.P. Moerman, K. Sergeant, G. Debyser, I. Timperman, B. Devreese, B. Samyn
Format: Article
Language:English
Published: Elsevier 2014-06-01
Series:EuPA Open Proteomics
Subjects:
C-terminal sequence analysis
Laboratory automation
Shewanella oneidensis MR-1
Proteomics
Online Access:http://www.sciencedirect.com/science/article/pii/S2212968514000257
Description
Summary:Experimental assignment of the protein termini remains essential to define the functional protein structure. Here, we report on the improvement of a proteomic C-terminal sequence analysis method. The approach aims to discriminate the C-terminal peptide in a CNBr-digest where Met-Xxx peptide bonds are cleaved in internal peptides ending at a homoserine lactone (hsl)-derivative. pH-dependent partial opening of the lactone ring results in the formation of doublets for all internal peptides. C-terminal peptides are distinguished as singlet peaks by MALDI-TOF MS and MS/MS is then used for their identification. We present a fully automated protocol established on a robotic liquid-handling station.
ISSN:2212-9685

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