Open AccessAffinity Chromatography of Collagen Glycosyltransferases on Collagen Linked to Agarose
Author(s) -
RISTELI Leila,
MYLLYLÄ Raili,
KIVIRIKKO Kari I.
Publication year - 1976
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1976.tb10649.x
Subject(s) - galactosyltransferase , cyanogen bromide , agarose , affinity chromatography , chemistry , glycosyltransferase , glucosyltransferase , sepharose , chromatography , biochemistry , size exclusion chromatography , ion chromatography , enzyme , peptide sequence , gene
Denatured citrate‐soluble collagen was coupled to agarose by the cyanogen bromide activation technique, and columns prepared from this material were studied for affinity chromatography of collagen glycosyltransferases. Both collagen glycosyltransferases became bound to the column, the degree of binding and the capacity of the column being higher with the glucosyltransferase than with the galactosyltransferase. The addition of Mn 2+ enhanced the binding, especially with the glucosyltransferase. The enzymes were eluted from the column with small peptides prepared from collagen, and they were separated from the peptides by gel filtration. With this procedure a collagen glucosyltransferase purification of about 5000‐fold and a collagen galactosyltransferase purification of about 1000‐fold was obtained from chick embryo extract by relatively simple steps.