Specific stimulation of alpha 2-6 sialyltransferase activity by a novel cytosolic factor from rat colon.
A factor present in the 100,000 g supernatant from the homogenate of rat colon stimulated the activity of purified Gal beta 1-4GLcNAc alpha 2,6 sialyltransferase [alpha 2-6ST(N)] from rat liver and alpha 2-6ST(N) from either liver microsomes or Golgi membrane. The stimulation of alpha 2-6ST(N) activity by the colon factor using protein acceptors was about four-fold and highly reproducible when the reaction product of the alpha 2-6ST(N) was assayed by either precipitation or affinity chromatography. In contrast, the colon factor did not stimulate the Gal beta 1-4GlcNAc alpha 2,3 sialyltransferase [alpha 2-3ST (N)], from rat jejunum microsomes or purified Gal beta 1-3GalNAc alpha 2,4 sialyltransferase [alpha 2-3ST (O)] from porcine liver, of purified beta 1,4 galactosyltransferase (GT) from bovine milk. In addition to rat colon, the 100,00 g supernatant from the homogenates of rat brain and kidney also stimulated the alpha 2-6ST(N) activity. The stimulation of alpha 2-6ST(N) by the colon factor resulted in a decrease in the Km (by about two-fold) and an increase in Vmax (about 2- to 3-fold) for desialylated alpha 1 acid glycoprotein and CMP-[14C]N-acetylneuraminic acid. The stimulation of alpha 2-6ST(N) activity by the colon factor was temperature dependent, protease sensitive and was inhibited by CTP, but did not need the presence of either metal ions or detergent. The cytosolic factor was partially purified by ion-exchange chromatography with the retention of the activator activity in the peaks containing low molecular weight proteins, but the activity was lost on attempts to further purification. A specific marked stimulation of the alpha 2-6ST(N) activity by cytosolic factors in certain tissues might suggest a physiological role for these factors in the regulation of alpha 2-5ST(N) activity.
Nagpurkar
A
A
Department of Biochemistry, Memorial University, St. John's, Newfoundland, Canada.
Hunt
D
D
Mookerjea
S
S
D000818:Animals; D001685:Biological Factors / Q000302:isolation & purification / Q000494:pharmacology*; D002240:Carbohydrate Sequence; D002417:Cattle; D003106:Colon / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004187:Disaccharides / Q000737:chemistry; D004789:Enzyme Activation; D006056:Golgi Apparatus / Q000201:enzymology; D066298:In Vitro Techniques; D007583:Jejunum / Q000201:enzymology; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008297:Male; D008861:Microsomes / Q000201:enzymology; D008862:Microsomes, Liver / Q000201:enzymology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012799:Sialyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D001685:Biological Factors; D004187:Disaccharides; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase