MUTATIONS OF HUMAN MYRISTOYL-COA-PROTEIN N-MYRISTOYLTRANSFERASE CAUSE TEMPERATURE-SENSITIVE MYRISTIC ACID AUXOTROPHY IN SACCHAROMYCES-CEREVISIAE
MUTATIONS OF HUMAN MYRISTOYL-COA-PROTEIN N-MYRISTOYLTRANSFERASE CAUSE TEMPERATURE-SENSITIVE MYRISTIC ACID AUXOTROPHY IN SACCHAROMYCES-CEREVISIAE
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DOI：
10.1073/pnas.89.9.4129
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发表时间：
1992-05-01
影响因子：
11.1
通讯作者：
GORDON, JI
中科院分区：
文献类型：
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作者：
DURONIO, RJ;REED, SI;GORDON, JI
我们通过补充酿酒酵母的 nmt1-181 突变（导致温度敏感的肉豆蔻酸营养缺陷型），分离出编码人肉豆蔻酰辅酶 A：蛋白 N-肉豆蔻酰转移酶（NMT，EC 2.3.1.97）的 cDNA。人NMT源自单拷贝基因，含有416个氨基酸，与酿酒酵母NMT（酵母NMT）有44%的一致性，并且可以补充nmt1无效突变的致死表型。根据在大肠杆菌中重建蛋白质 N-肉豆蔻酰化的共表达系统判断，人和酵母 NMT 具有重叠但不同的蛋白质底物特异性。两种酶都含有距 C 末端 5 个残基的甘氨酸。这些直系同源 NMT 中的 Gly --> Asp 或 Lys 诱变导致其在大肠杆菌中的活性显着降低，并且在酿酒酵母中导致温度敏感的肉豆蔻酸营养缺陷型。这些结果表明体内高度保守的结构-功能关系，并强调了这些功能测定对于识别哺乳动物系统中调节蛋白质 N-肉豆蔻酰化的因子的有用性。
We have isolated cDNAs encoding human myristoyl-CoA:protein N-myristoyltransferase (NMT, EC 2.3.1.97) by complementing the nmt1-181 mutation of Saccharomyces cerevisiae, which causes temperature-sensitive myristic acid auxotrophy. Human NMT is derived from a single-copy gene, contains 416 amino acids, is 44% identical to S. cerevisiae NMT (yeast NMT), and can complement the lethal phenotype of an nmt1 null mutation. Human and yeast NMTs have overlapping yet distinct protein substrate specificities as judged by a coexpression system that reconstitutes protein N-myristoylation in Escherichia coli. Both enzymes contain a glycine five residues from the C terminus. Gly --> Asp or Lys mutagenesis in these orthologous NMTs produces marked reductions in their activities in E. coli as well as temperature-sensitive myristic acid auxotrophy in S. cerevisiae. These results indicate highly conserved structure-function relationships in vivo and underscore the usefulness of these functional assays for identifying factors that regulate protein N-myristoylation in mammalian systems.