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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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
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spelling doaj-0cc00214de6c41e798e663b20167a65f2020-11-24T23:55:12ZengElsevierEuPA Open Proteomics2212-96852014-06-013C25026110.1016/j.euprot.2014.03.004Automation of C-terminal sequence analysis of 2D-PAGE separated proteinsP.P. Moerman0K. Sergeant1G. Debyser2I. Timperman3B. Devreese4B. Samyn5Ghent University, Department Biochemistry and Microbiology, Laboratory Protein Biochemistry and Biomolecular Engineering – L-ProBE, KL Ledeganckstraat 35, B-9000 Ghent, BelgiumCentre de Recherche Public – Gabriel Lippmann, Department ‘Environment and Agro-biotechnologies’ (EVA), CRP-GL, Rue du Brill 41, 4422 Belvaux, LuxembourgGhent University, Department Biochemistry and Microbiology, Laboratory Protein Biochemistry and Biomolecular Engineering – L-ProBE, KL Ledeganckstraat 35, B-9000 Ghent, BelgiumGhent University, Department Biochemistry and Microbiology, Laboratory Protein Biochemistry and Biomolecular Engineering – L-ProBE, KL Ledeganckstraat 35, B-9000 Ghent, BelgiumGhent University, Department Biochemistry and Microbiology, Laboratory Protein Biochemistry and Biomolecular Engineering – L-ProBE, KL Ledeganckstraat 35, B-9000 Ghent, BelgiumGhent University, Department Biochemistry and Microbiology, Laboratory Protein Biochemistry and Biomolecular Engineering – L-ProBE, KL Ledeganckstraat 35, B-9000 Ghent, BelgiumExperimental 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.http://www.sciencedirect.com/science/article/pii/S2212968514000257C-terminal sequence analysisLaboratory automationShewanella oneidensis MR-1Proteomics
collection DOAJ
language English
format Article
sources DOAJ
author P.P. Moerman
K. Sergeant
G. Debyser
I. Timperman
B. Devreese
B. Samyn
spellingShingle P.P. Moerman
K. Sergeant
G. Debyser
I. Timperman
B. Devreese
B. Samyn
Automation of C-terminal sequence analysis of 2D-PAGE separated proteins
EuPA Open Proteomics
C-terminal sequence analysis
Laboratory automation
Shewanella oneidensis MR-1
Proteomics
author_facet P.P. Moerman
K. Sergeant
G. Debyser
I. Timperman
B. Devreese
B. Samyn
author_sort P.P. Moerman
title Automation of C-terminal sequence analysis of 2D-PAGE separated proteins
title_short Automation of C-terminal sequence analysis of 2D-PAGE separated proteins
title_full Automation of C-terminal sequence analysis of 2D-PAGE separated proteins
title_fullStr Automation of C-terminal sequence analysis of 2D-PAGE separated proteins
title_full_unstemmed Automation of C-terminal sequence analysis of 2D-PAGE separated proteins
title_sort automation of c-terminal sequence analysis of 2d-page separated proteins
publisher Elsevier
series EuPA Open Proteomics
issn 2212-9685
publishDate 2014-06-01
description 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.
topic C-terminal sequence analysis
Laboratory automation
Shewanella oneidensis MR-1
Proteomics
url http://www.sciencedirect.com/science/article/pii/S2212968514000257
work_keys_str_mv AT ppmoerman automationofcterminalsequenceanalysisof2dpageseparatedproteins
AT ksergeant automationofcterminalsequenceanalysisof2dpageseparatedproteins
AT gdebyser automationofcterminalsequenceanalysisof2dpageseparatedproteins
AT itimperman automationofcterminalsequenceanalysisof2dpageseparatedproteins
AT bdevreese automationofcterminalsequenceanalysisof2dpageseparatedproteins
AT bsamyn automationofcterminalsequenceanalysisof2dpageseparatedproteins
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