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Biosynthesis of bloodgroup I and i antigens. A sensitive and specific assay of UDP‐GlcNAc:β‐galactoside β1 → 3‐N‐acetylglucosaminyltransferase activity in hematopoietic cells by HPLC
Author(s) -
Koenderman Anky H. L.,
Loppen Piet L.,
Marinus Lian A. M.,
van den Eijnden Dirk H.
Publication year - 1986
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130010304
Subject(s) - chemistry , high performance liquid chromatography , substrate (aquarium) , in vitro , enzyme , biochemistry , glycosyltransferase , transferase , lactose , haematopoiesis , chromatography , cell culture , biosynthesis , stem cell , biology , microbiology and biotechnology , ecology , genetics
A sensitive HPLC method for the assay of UDP‐GlcNAc:β‐galactoside β1 → 3‐ N ‐acetylglucosaminyltransferase activity was developed. Using lactose as an acceptor, the formation of the product GlcNAcβ1 → 3Galβ1 → 4Glc can be determined without interference by substrates resulting from enzymatic and chemical breakdown of the donor substrate UDP‐GlcNAc. The method is very specific since products of other transferase reactions, which potentially may be formed in the incubations in vitro , elute at positions different from that of GlcNAcβ1 → 3Galβ1 → 4Glc. By use of this assay method it could be demonstrated that normal and malignant hematopoietic cells and cell‐lines, with the exception of erythrocytes and reticulocytes, contain β1 → 3‐ N ‐acetylglucosaminyltransferase activity.