Discrimination of glycoproteins from unglycosylated proteins in capillary electrophoresis: Two-color LIF detection coupled with post-column derivatization

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

Glycosylation is one of the most important posttranslational modifications (PTMs) which lead to the functionalization of proteins. Here, we describe one method for discriminating glycosylated proteins from unglycosylated ones in their mixture sample by capillary electrophoretic separation and two-color laserinduced fluorescence detection coupled with post-column derivatization. Two lasers emitting at 450 and 532 nm permit the detection of amino groups of proteins derivatized by naphthalene-2,3-dicarboxaldehyde and a fluorescently labeled lectin, tetramethylrhodamine-labeled concanavalin A (Rh-Con A), respectively. When a protein mixture react with Rh-Con A, the glycoproteins bound with Rh-Con A exhibit signals at the same migration time in two electropherograms obtained by 450-and 532-nm lasers whereas unbound proteins show a signal only in the electropherogram of the 450-nm laser. So, when one protein is glycosylated it is detected at the same migration time in the electropherograms obtained by two lasers.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages11-23
Number of pages13
Volume1466
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1466
ISSN (Print)1064-3745

Fingerprint

Capillary Electrophoresis
Glycoproteins
Color
Lasers
Proteins
Post Translational Protein Processing
Concanavalin A
Glycosylation
Lectins
Fluorescence

Keywords

  • Capillary electrophoresis
  • Glycoprotein
  • Postcolumn derivatization
  • Two-color laserinduced fluorescence

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Discrimination of glycoproteins from unglycosylated proteins in capillary electrophoresis : Two-color LIF detection coupled with post-column derivatization. / Kaneta, Takashi.

Methods in Molecular Biology. Vol. 1466 Humana Press Inc., 2016. p. 11-23 (Methods in Molecular Biology; Vol. 1466).

Research output: Chapter in Book/Report/Conference proceedingChapter

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AB - Glycosylation is one of the most important posttranslational modifications (PTMs) which lead to the functionalization of proteins. Here, we describe one method for discriminating glycosylated proteins from unglycosylated ones in their mixture sample by capillary electrophoretic separation and two-color laserinduced fluorescence detection coupled with post-column derivatization. Two lasers emitting at 450 and 532 nm permit the detection of amino groups of proteins derivatized by naphthalene-2,3-dicarboxaldehyde and a fluorescently labeled lectin, tetramethylrhodamine-labeled concanavalin A (Rh-Con A), respectively. When a protein mixture react with Rh-Con A, the glycoproteins bound with Rh-Con A exhibit signals at the same migration time in two electropherograms obtained by 450-and 532-nm lasers whereas unbound proteins show a signal only in the electropherogram of the 450-nm laser. So, when one protein is glycosylated it is detected at the same migration time in the electropherograms obtained by two lasers.

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KW - Two-color laserinduced fluorescence

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