Identification of rat alpha1 macroglobulin as an inhibitor of rat Galbeta1-4GlcNAc alpha2-6 sialyltransferase.
Rat serum was found to contain an inhibitor of pure alpha2-6 sialyltransferase (EC 2.4.99.1). The inhibitor was a high Mr protein isolated by molecular filtration on Sephadex G100, followed by anion exchange chromatography on Sephadex DEAE A25, then separation on Sepharose CL 4B, and finally by isoelectric focusing. Electrophoretic separation and subsequent N-terminal sequence analysis of the active inhibitor preparation showed the presence of two protein components, identified as rat C-reactive protein, and rat alpha1 macroglobulin. Pure rat CRP did not inhibit alpha2-6 sialyltransferase. Treatment of the inhibitor preparations with monospecific antibodies against rat alpha1 macroglobulin blocked inhibitory activity in a dose-dependent manner. The results present strong evidence that alpha1 macroglobulin is capable of acting as an inhibitor of alpha2-6 sialyltransferase. No inhibition of galactosyltransferase (EC 2.4.1.38) could be detected, indicating that the interaction with alpha1 macroglobulin may have specificity among the glycosyltransferases. The entrapment of alpha2-6 sialyltransferase by alpha1 macroglobulin presented here occurs in vitro and will require further in vivo investigations to determine the precise physiological significance. Independent of the physiologic significance is the finding that this interaction occurs in vitro, which, pending elucidation of the precise mechanism and specificity, may provide a new tool for investigations into the functional significance of sialylation, and potentially for use or design of new inhibitors of therapeutic value in physiologic conditions involving altered levels of sialylation.
Harder
P G
PG
Department of Chemistry, University of Manitoba, Winnipeg, Canada.
Jamieson
J C
JC
D000595:Amino Acid Sequence; D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D004791:Enzyme Inhibitors / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats; D012799:Sialyltransferases / Q000037:antagonists & inhibitors*; D013379:Substrate Specificity; D000511:alpha-Macroglobulins / Q000302:isolation & purification / Q000378:metabolism*; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D004791:Enzyme Inhibitors; D000511:alpha-Macroglobulins; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase