STRUCTURAL CHARACTERIZATION OF GLYCOPROTEIN CARBOHYDRATE CHAINS BY USING DIGOXIGENIN-LABELED LECTINS ON BLOTS

被引:146
作者
HASELBECK, A
SCHICKANEDER, E
VONDERELTZ, H
HOSEL, W
机构
[1] Boehringer Mannheim GmbH, Biochemica Research Center
关键词
D O I
10.1016/0003-2697(90)90381-I
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The carbohydrate structures of blotted glycoproteins can be analyzed by probing them with lectins. Here we describe a method where lectins conjugated with digoxigenin are used in combination with an anti-digoxigenin antibody AP conjugate as a very sensitive detection system for this type of analysis. The specificity of the lectins used, and the sensitivity of the detection system, provide valuable conclusions on the glycan structures. Only small amounts of glycoproteins are required for the analysis. The binding specificity of a set of lectins is demonstrated with various glycoproteins of defined carbohydrate structure. The application of these labeled lectins in combination with specific glycosidases for the characterization of the carbohydrate chains of recombinant tissue plasminogen activator and erythropoietin is presented. © 1990.
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页码:25 / 30
页数:6
相关论文
共 32 条
[1]  
ANIMASHAUN T, 1989, J BIOL CHEM, V264, P4657
[2]  
BURNETTE WN, 1981, ANAL BIOCHEM, V112, P195, DOI 10.1016/0003-2697(81)90281-5
[4]   CARBOHYDRATE BINDING-STUDIES ON THE LECTIN FROM DATURA-STRAMONIUM SEEDS [J].
CROWLEY, JF ;
GOLDSTEIN, IJ ;
ARNARP, J ;
LONNGREN, J .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1984, 231 (02) :524-533
[5]  
DEBRAY H, 1981, EUR J BIOCHEM, V117, P41
[6]   ALEURIA-AURANTIA AGGLUTININ - A NEW ISOLATION PROCEDURE AND FURTHER STUDY OF ITS SPECIFICITY TOWARDS VARIOUS GLYCOPEPTIDES AND OLIGOSACCHARIDES [J].
DEBRAY, H ;
MONTREUIL, J .
CARBOHYDRATE RESEARCH, 1989, 185 (01) :15-26
[7]   USE OF LECTINS FOR DETECTION OF ELECTROPHORETICALLY SEPARATED GLYCOPROTEINS TRANSFERRED ONTO NITROCELLULOSE SHEETS [J].
GLASS, WF ;
BRIGGS, RC ;
HNILICA, LS .
ANALYTICAL BIOCHEMISTRY, 1981, 115 (01) :219-224
[8]  
GREEN ED, 1988, J BIOL CHEM, V263, P18253
[9]   DESCRIPTION AND APPLICATION OF AN IMMUNOLOGICAL DETECTION SYSTEM FOR ANALYZING GLYCOPROTEINS ON BLOTS [J].
HASELBECK, A ;
HOSEL, W .
GLYCOCONJUGATE JOURNAL, 1990, 7 (01) :63-74
[10]   CARBOHYDRATE CHAINS ON YEAST CARBOXYPEPTIDASE-Y ARE PHOSPHORYLATED [J].
HASHIMOTO, C ;
COHEN, RE ;
ZHANG, WJ ;
BALLOU, CE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1981, 78 (04) :2244-2248