TY - JOUR
T1 - Collection of α1-acid glycoprotein molecular species by capillary electrophoresis and the analysis of their molecular masses and carbohydrate chains
T2 - Basic studies on the analysis of glycoprotein glycoforms
AU - Sei, Kazuo
AU - Nakano, Miyako
AU - Kinoshita, Mitsuhiro
AU - Masuko, Takashi
AU - Kakehi, Kazuaki
PY - 2002/6/7
Y1 - 2002/6/7
N2 - A highly heterogeneous glycoprotein, α1-acid glycoprotein, was resolved into their glycoforms by capillary electrophoresis using a surface-modified capillary in 20 mM acetate buffer (pH 4.2) containing 0.5% (w/v) hydroxypropylmethylcellulose. We collected the fractions containing each glycoform as nearly pure state by capillary electrophoresis, and examined the molecular masses of these glycoforms by matrix assisted laser desorption time-of-flight mass spectrometry. We also analyzed carbohydrate chains after releasing them with N-glycosidase F followed by fluorescent labeling with 8-aminopyrene-1,3,6-trisulfonate. We found that the separation of glycoforms was mostly due to the presence of multiantennary carbohydrate chains. We propose that the present technique is useful for the analysis of post translational modification of proteins with carbohydrate chains.
AB - A highly heterogeneous glycoprotein, α1-acid glycoprotein, was resolved into their glycoforms by capillary electrophoresis using a surface-modified capillary in 20 mM acetate buffer (pH 4.2) containing 0.5% (w/v) hydroxypropylmethylcellulose. We collected the fractions containing each glycoform as nearly pure state by capillary electrophoresis, and examined the molecular masses of these glycoforms by matrix assisted laser desorption time-of-flight mass spectrometry. We also analyzed carbohydrate chains after releasing them with N-glycosidase F followed by fluorescent labeling with 8-aminopyrene-1,3,6-trisulfonate. We found that the separation of glycoforms was mostly due to the presence of multiantennary carbohydrate chains. We propose that the present technique is useful for the analysis of post translational modification of proteins with carbohydrate chains.
KW - Alpha1-acid glycoprotein
KW - Carbohydrates
KW - Glycoproteins
KW - Orosomucoid
KW - Proteins
UR - http://www.scopus.com/inward/record.url?scp=0037035838&partnerID=8YFLogxK
U2 - 10.1016/S0021-9673(02)00353-9
DO - 10.1016/S0021-9673(02)00353-9
M3 - Article
C2 - 12134825
AN - SCOPUS:0037035838
SN - 0021-9673
VL - 958
SP - 273
EP - 281
JO - Journal of Chromatography A
JF - Journal of Chromatography A
IS - 1-2
ER -