Special considerations in the purification of the GM3 ganglioside-forming enzyme, CMP-sialic acid:lactosylceramide alpha 2-3 sialyltransferase (SAT-1): effects of protease inhibitors on rat hepatic SAT-1 activity.
Special considerations in the purification of the GM3 ganglioside-forming enzyme, CMP-sialic acid:lactosylceramide alpha 2-3 sialyltransferase (SAT-1): effects of protease inhibitors on rat hepatic SAT-1 activity.
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GM3 神经节苷脂形成酶 CMP-唾液酸：乳糖神经酰胺 α 2-3 唾液酸转移酶 (SAT-1) 纯化中的特殊考虑因素：蛋白酶抑制剂对大鼠肝 SAT-1 活性的影响。
DOI：
10.1016/s0006-291x(05)81238-0
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发表时间：
1991
影响因子：
3.1
通讯作者：
Sweeley,CC
中科院分区：
文献类型：
--
作者：
Melkerson-Watson,LJ;Sweeley,CC
内源蛋白水解活性的共纯化被认为是唾液酸转移酶大小异质性的原因。本文报道了各种蛋白酶抑制剂、巯基还原剂和抗微生物剂对 SAT-1 活性的影响的结果。在免疫亲和纯化的大鼠肝脏 SAT-1 中添加蛋白酶抑制剂会显着影响其活性。除 PMSF 外，所有检测的蛋白酶抑制剂均抑制纯化的酶。最具抑制性的是半胱氨酸（硫醇）蛋白酶抑制剂。当研究这些抑制剂对富含高尔基体的微粒体中 SAT-1 活性的影响时，这种效果并不那么引人注目，尽管半胱氨酸蛋白酶抑制剂的抑制作用最大。在这两种情况下都发现了一个显着的效果，那就是在 β-巯基乙醇存在的情况下 SAT1 活性明显激活。
Co-purification of an endogenous proteolytic activity has been proposed as the cause for the size heterogeneity of sialyltransferases. Reported herein are results on the effects of various protease inhibitors, sulfhydryl-reducing agents and antimicrobial agents on SAT-1 activity. Addition of protease inhibitors to immunoaffinity-purified rat liver SAT-1 dramatically affects its activity. All protease inhibitors examined, with the exception of PMSF, inhibited the purified enzyme. The most inhibitory were the cysteine (thiol) protease inhibitors. This effect is less spectacular when the effect of these inhibitors was studied on SAT-1 activity in Golgi-enriched microsomes, although the inhibition was greatest by the cysteine protease inhibitors. One dramatic effect, found in both cases, was the apparent activation of SAT1 activity in the presence of β-mercaptoethanol.