Bergeron J J, Rachubinski R A, Sikstrom R A, Posner B I, Paiement J
J Cell Biol. 1982 Jan;92(1):139-46. doi: 10.1083/jcb.92.1.139.
The distribution of galactosyl transferase was studied using trans and cis Golgi fractions isolated by a modification of the Ehrenreich et al. procedure (1973. J. Cell Biol. 59:45-72) as well as an intact Golgi fraction isolated by a new one-step procedure. Two methods of assay were used. The first method analyzed the ability of Golgi fractions to transfer galactose (from uridine diphosphogalactose [UDP-gal] substrate) to the defined exogenous acceptor ovomucoid. The second method assessed the transfer of galactose from UDP-gal substrate to endogenous acceptors (endogenous glycosylation). The trans Golgi fraction (Golgi light) was highly active by the first method but revealed only low activity by the second method. Golgi fractions enriched in central and cis elements (the Golgi intermediate, heavy and especially the intact Golgi fraction) were highly active in both methods of assay. The endogenous glycosylation approach was validated by gel fluorography of the endogenous acceptors. For all Golgi fractions, transfer of galactose was revealed to secretory glycopeptides. It is concluded that galactosyl transferase activity in vivo occurs primarily in central and cis Golgi elements but not trans Golgi vesicles.
利用经改良的埃伦赖希等人（1973年，《细胞生物学杂志》59卷：45 - 72页）的方法分离得到的反式和顺式高尔基体组分，以及通过一种新的一步法分离得到的完整高尔基体组分，研究了半乳糖基转移酶的分布情况。采用了两种测定方法。第一种方法分析高尔基体组分将半乳糖（来自尿苷二磷酸半乳糖[UDP - gal]底物）转移至特定外源受体卵类黏蛋白的能力。第二种方法评估半乳糖从UDP - gal底物向内源受体的转移（内源糖基化）。反式高尔基体组分（高尔基体轻组分）通过第一种方法表现出高活性，但通过第二种方法仅显示出低活性。富含中央和顺式元件的高尔基体组分（高尔基体中间组分、重组分，尤其是完整高尔基体组分）在两种测定方法中均表现出高活性。通过对内源受体进行凝胶荧光成像验证了内源糖基化方法。对于所有高尔基体组分，半乳糖的转移均显示于分泌型糖肽中。得出的结论是，体内半乳糖基转移酶活性主要发生在中央和顺式高尔基体元件中，而非反式高尔基体小泡中。
