title: Discrimination of Glycoproteins from Unglycosylated Proteins in Capillary Electrophoresis: Two-Color LIF Detection Coupled with Post-column Derivatization.
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 laser-induced 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.
authors.affiliation: Department of Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushimanaka 3-1-1, Kita-ku, Okayama-shi, 700-8530, Japan. kaneta@okayama-u.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kaneta
mesh_terms: D003208:Concanavalin A / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D006023:Glycoproteins / Q000302:isolation & purification*; D009281:Naphthalenes / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry; D013050:Spectrometry, Fluorescence
chemical_list: D006023:Glycoproteins; D009281:Naphthalenes; D012235:Rhodamines; D003208:Concanavalin A; C005358:tetramethylrhodamine; C027717:2,3-naphthalenedicarboxaldehyde
keywords: Capillary electrophoresis; Glycoprotein; Postcolumn derivatization; Two-color laser-induced fluorescence