Glycoproteins with lectins

B13. B0g-Hansen, T. C., Affinity electrophoresis. Quantification and characterisation of glycoproteins with lectins. In Electrophoresis 79 . (B. J. Radola, ed.), pp. 193-202. Walter de Gruyter, Berlin, 1980. 

For the characterization of glycoproteins with lectins, the sample is run on an SDS gel with an approximate 10-cm-wide pocket and then blotted. The blot (on PVDF membrane) is stained on protein and the approximately 10-cm-long protein band is cut crosswise into approximately 20 narrow strips. After blocking (e.g., with 2% polyvinylpyrrolidon 360 in 50 mM Tris-Cl pH 7.5 and 0.5 M NaCl), the strips are incubated in twos with a labeled lectin (Table 9.1) with or without inhibiting sugar, and are then developed. 

B. Hydroxyproline-Rich Glycoproteins With Lectin-Like Properties.. 234... 

Rohnnger, R. and Holden, D W (1985) Protein blotting detection of protein with colloidal gold, and of glycoproteins and lectins with biotin-conjugated and enzyme probes Anal Biochem. 144, 118-127. 

The seeds of Griffonia (Bandeiraea) simplicifolia contain a second lectin - GS II. It is a tetramer composed of four apparently identical subunits (A/r = 30000) with an aggregate, Mr = 113 000 Da by gel filtration [142]. The lectin is a glycoprotein with three cysteine residues per subunit a disulfide bond links two subunits [142], Each subunit contains a binding site for GlcNAc with ATa= 1.3x 10 M" at 37"C [143]. 

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