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 language | English |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 11-23 |
| Number of pages | 13 |
| Volume | 1466 |
| DOIs | |
| Publication status | Published - 2016 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1466 |
| ISSN (Print) | 1064-3745 |
Fingerprint
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 proceeding › Chapter
}
TY - CHAP
T1 - Discrimination of glycoproteins from unglycosylated proteins in capillary electrophoresis
T2 - Two-color LIF detection coupled with post-column derivatization
AU - Kaneta, Takashi
PY - 2016
Y1 - 2016
N2 - 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.
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.
KW - Capillary electrophoresis
KW - Glycoprotein
KW - Postcolumn derivatization
KW - Two-color laserinduced fluorescence
UR - http://www.scopus.com/inward/record.url?scp=84990222543&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84990222543&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-4014-1_2
DO - 10.1007/978-1-4939-4014-1_2
M3 - Chapter
C2 - 27473478
AN - SCOPUS:84990222543
VL - 1466
T3 - Methods in Molecular Biology
SP - 11
EP - 23
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -