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Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes

Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes

Published on: November 26, 2008

Pyrylium-based dye and charge tagging in proteomics.

Malte Bayer¹, Simone König¹

¹Core Unit Proteomics, Interdisciplinary Center for Clinical Research, University of Münster, Münster, Germany.

Electrophoresis

|August 2, 2016

Summary

The pyrylium dye Py-1 selectively labels lysine residues and N-termini in peptides, offering advantages for fluorescence and charge tagging in proteomics research.

Keywords:

Charge tag Fluorescent dye Gel electrophoresis MS Oxonium ion Proteomics Pyrylium cation

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Area of Science:

Chemical Biology
Proteomics
Analytical Chemistry

Background:

Pyrylium dyes offer advantages for protein labeling, including rapid procedures.
Traditional N-hydroxysuccinimidyl ester chemistry is commonly used for labeling.

Purpose of the Study:

Investigate the specificity and side products of pyrylium dye Py-1 labeling.
Evaluate Py-1 for fluorescence, N-terminal charge, and chromophore tagging.

Main Methods:

Liquid chromatography coupled to mass spectrometry (LC-MS) was used.
Peptides with varying lysine and cysteine residues were analyzed.
Gas phase fragmentation was employed to identify labeling sites.

Main Results:

Py-1 labeling of lysine residues and N-termini was confirmed with yields >30%.
No evidence of cysteine labeling was found.
Side products from rearrangements with acidic amino acids were detected.

Conclusions:

Py-1 is a selective reagent for lysine and N-terminal labeling in peptides.
Py-1 serves as a fluorogenic label, N-terminal charge tag, and chromophore tag.
The dye shows potential as an alternative to existing labeling chemistries.