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GM3神经节苷脂形成酶CMP-唾液酸:乳糖基神经酰胺α2-3唾液酸转移酶(SAT-1)纯化中的特殊考虑因素:用月桂基二甲基氧化胺溶解SAT-1

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.

作者信息

Melkerson-Watson L J, Sweeley C C

机构信息

Department of Biochemistry, Michigan State University, East Lansing 48824.

出版信息

Biochem Biophys Res Commun. 1990 Oct 15;172(1):165-71. doi: 10.1016/s0006-291x(05)80188-3.

Abstract

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.

摘要

月桂基二甲基氧化胺(LDAO)被用于纯化GM3神经节苷脂形成酶,即CMP-唾液酸:乳糖基神经酰胺α2-3唾液酸转移酶(SAT-1)(4)。在溶解和稳定这种唾液酸转移酶方面,这种去污剂比通常使用的Triton去污剂具有优势。通过二维凝胶电泳测定,用几种此类去污剂从大鼠肝脏高尔基体中溶解出的粗蛋白组分在组成上非常相似。然而,LDAO似乎能激活并稳定SAT-1的活性。SAT-1的激活可能涉及LDAO和磷脂酰胆碱结构相似的疏水部分和季铵基团。

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