N-acetylglucosaminyltransferase III, IV and V activities in Novikoff ascites tumour cells, mouse lymphoma cells and hen oviduct. Application of a sensitive and specific assay by use of high-performance liquid chromatography.
A specific and fast method for the determination of N-acetylglucosaminyltransferase III, IV and V activity in one assay is described. The method is based on the separation by HPLC of the three transferase products formed from the common acceptor oligosaccharide substrate GlcNAc beta 1----2Man alpha 1----3(GlcNAc beta 1----2Man alpha 1---- 6)Man beta 1----4GlcNAc. Assays are not interfered with by substances that result from enzymatic or chemical breakdown of the donor substrate UDP-[14C]GlcNAc. Using this assay system N-acetylglucosaminyltransferase III, IV and V activities were estimated in Novikoff ascites tumour cells, mouse lymphoma BW 5147 cells and hen oviduct.
Koenderman
A H
AH
Department of Medical Chemistry, Vrije Universiteit, Amsterdam, The Netherlands.
Koppen
P L
PL
Koeleman
C A
CA
van den Eijnden
D H
DH
D000818:Animals; D002460:Cell Line; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005964:Glucosyltransferases / Q000302:isolation & purification*; D008223:Lymphoma / Q000201:enzymology; D017351:N-Acetylglucosaminyltransferases; D009374:Neoplasms, Experimental / Q000201:enzymology*; D009844:Oligosaccharides / Q000096:biosynthesis; D010057:Oviducts / Q000201:enzymology* / Q000378:metabolism; D051381:Rats; D014166:Transferases / Q000302:isolation & purification
D009844:Oligosaccharides; D014166:Transferases; D005964:Glucosyltransferases; D017351:N-Acetylglucosaminyltransferases; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase; C038111:alpha-1,3-mannosylglycoprotein beta-1,4-N-acetylglucosaminyltransferase; C052254:alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase