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一种用于检测负责将肉豆蔻酸连接到蛋白质N端甘氨酸残基的酶(肉豆蔻酰辅酶A:甘氨酰肽N-肉豆蔻酰转移酶)的简化检测方法。

A simplified assay for the enzyme responsible for the attachment of myristic acid to the N-terminal glycine residue of proteins, myristoyl-CoA: glycylpeptide N-myristoyltransferase.

作者信息

McIlhinney R A, McGlone K

机构信息

Department of Pharmacology, University of Oxford, U.K.

出版信息

Biochem J. 1989 Oct 15;263(2):387-91. doi: 10.1042/bj2630387.

Abstract

A greatly simplified assay for myristoyl-CoA:glycylpeptide N-myristoyltransferase (NMT) activity is described. The assay is based on the differential solubility of the acyl-peptides produced as a consequence of the NMT activity and yields results comparable with those obtained with the original assay described by Towler & Glaser [(1986) Proc. Natl. Acad. Sci. U.S.A. 83, 2812-2816], which requires h.p.l.c. to determine the production of the acyl-peptides. The use of the revised assay in the preliminary steps of the purification of rat brain NMT is described, and its use in determining the fatty acid-specificity of the enzyme is illustrated. The results are shown to be comparable with those obtained with the h.p.l.c.-based assay.

摘要

本文描述了一种用于测定肉豆蔻酰辅酶A:甘氨酰肽N-肉豆蔻酰转移酶(NMT)活性的大大简化的检测方法。该检测方法基于因NMT活性产生的酰基肽的不同溶解度,其产生的结果与Towler和Glaser所描述的原始检测方法[(1986)美国国家科学院院刊83, 2812 - 2816]所得结果相当,原始方法需要高效液相色谱法来测定酰基肽的产生。本文描述了该改进检测方法在大鼠脑NMT纯化初步步骤中的应用,并举例说明了其在确定该酶的脂肪酸特异性方面的应用。结果表明,该方法所得结果与基于高效液相色谱法的检测结果相当。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4749/1133441/5b355967e0cf/biochemj00197-0080-a.jpg

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