Special considerations in the purification of the GM3 ganglioside forming enzyme, CMP-sialic acid:lactosylceramide alpha 2-3 sialyltransferase (SAT-1): solubilization of SAT-1 with lauryldimethylamine oxide.
Lauryldimethylamine oxide (LDAO) was employed in the purification of the GM3 ganglioside forming enzyme, CMP-sialic acid:lactosylceramide alpha 2-3 sialyltransferase (SAT-1) (4). This detergent has advantages over the typically employed Triton detergents in the solubilization and stabilization of this sialyltransferase. Crude protein fractions solubilized from rat liver Golgi by several such detergents are very similar in composition as determined by two-dimensional gel electrophoresis. However, LDAO appears to activate and stabilize SAT-1 activity. It is possible that SAT-1 activation involves the structurally similar hydrophobic moieties and quaternary amino groups of LDAO and phosphatidylcholine.
Melkerson-Watson
L J
LJ
Department of Biochemistry, Michigan State University, East Lansing 48824.
Sweeley
C C
CC
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003902:Detergents / Q000494:pharmacology; D004123:Dimethylamines; D015180:Electrophoresis, Gel, Two-Dimensional; D005679:G(M3) Ganglioside / Q000096:biosynthesis*; D006056:Golgi Apparatus / Q000201:enzymology*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D051381:Rats; D012799:Sialyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
D003902:Detergents; D004123:Dimethylamines; D005679:G(M3) Ganglioside; C014518:dodecyldimethylamine oxide; D012799:Sialyltransferases; C021343:haematoside synthetase