ISSN:
1573-4986
Keywords:
sialyltransferase
;
colostrum
;
CDP-ethanolamine-(aminocaproyl)-agarose
Source:
Springer Online Journal Archives 1860-2000
Topics:
Chemistry and Pharmacology
Notes:
Abstract Cytidine-5′-monophospho-N-acetylneuraminic acid:β-galactoside α2-6sialyltransferase was purified from bovine colostrum by two sequential affinity chromatography steps on CDP-ethanolamine-Sepharose and CDP-ethanolamine-(N-caproylamino-)-Sepharose, respectively. While the conditions for elution were those of Paulsonet al. [J Biol Chem (1977) 252:3256–62], the ligand of the second affinity column was coupled to Sepharose by using 6-aminocaproic acid as linker. The ease of this procedure allows rapid synthesis of bulk quantities of ligand. Highly purified preparations of sialyltransferase were obtained which moved on gradient gel electrophoresis as a single band of 76 kDa and on dodecylsulphate electrophoresis as a single band of 54 kDa. The product of the reaction between lactose and CMP-N-acetylneuraminic acid catalyzed by the purified sialyltransferase was identified by high-resolution 500 MHz1H-NMR spectroscopy as Neu5Acα2-6Galβ1-4Glc.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01213727
