Trinchera M, Ghidoni R
Department of Medical Chemistry and Biochemistry, Medical School, University of Milan, Italy.
J Biol Chem. 1989 Sep 25;264(27):15766-9.
A highly purified Golgi preparation from rat liver was fractionated on a sucrose density gradient and the activity of two sialyltransferases, CMP-NeuAc: Gal beta 1----4Glc-Cer (lactosylceramide) alpha-2----3sialyltransferase; Sat-1), and CMP-NeuAc:Gal beta 1----3GalNAc beta 1----4(NeuAc alpha 2----3) Gal beta 1----4Glc-Cer (GM1 ganglioside) alpha 2----3sialyltransferase; SAT-4), involved in the biosynthesis of gangliosides were assayed in the collected fractions. These two activities were recovered in different regions of the gradient; SAT-1 was found in a more dense region than SAT-4. This distribution coincided with that of two N-Asn linked oligosaccharide processing enzymes (UDP-GlcNAc:lysosomal enzyme precursor GlcNAc-1-phosphotransferase and UDP-Gal:ovalbumin galactosyltransferase), assumed as putative markers of cis- and trans-Golgi cisternae, respectively. These findings are consistent with the assembly of ganglioside oligosaccharide chains occurring in different sub-Golgi compartments.
对从大鼠肝脏中提取的高度纯化的高尔基体提取物进行蔗糖密度梯度分级分离,并对收集的各组分中参与神经节苷脂生物合成的两种唾液酸转移酶的活性进行测定,这两种酶分别是CMP-神经氨酸:Galβ1----4Glc-Cer(乳糖基神经酰胺)α-2----3唾液酸转移酶(Sat-1)和CMP-神经氨酸:Galβ1----3GalNAcβ1----4(NeuAcα2----3)Galβ1----4Glc-Cer(GM1神经节苷脂)α2----3唾液酸转移酶(SAT-4)。这两种活性在梯度的不同区域中回收;发现Sat-1所在区域比SAT-4所在区域密度更大。这种分布与两种N-Asn连接的寡糖加工酶(UDP-GlcNAc:溶酶体酶前体GlcNAc-1-磷酸转移酶和UDP-Gal:卵清蛋白半乳糖基转移酶)的分布一致,这两种酶分别被认为是顺式和反式高尔基体潴泡的假定标志物。这些发现与神经节苷脂寡糖链在不同高尔基体亚区室中的组装情况一致。