Eukaryotic initiation factor 4D, the hypusine-containing protein, is conserved among eukaryotes.
Eukaryotic initiation factor 4D, the hypusine-containing protein, is conserved among eukaryotes.
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DOI：
10.1016/s0021-9258(18)49296-4
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发表时间：
1987-12
期刊：
影响因子：
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通讯作者：
Edys D. Gordon;Rene MorallII;Stephen;-C.;Meredith;Chung Lee;Susan;-L.;Lindquistss
中科院分区：
文献类型：
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作者：
Edys D. Gordon;Rene MorallII;Stephen;-C.;Meredith;Chung Lee;Susan;-L.;Lindquistss
当哺乳动物细胞在含有[3H]亚精胺的培养基中生长时，一个与真核起始因子4D相同的单一主要氚化蛋白被标记。这种蛋白质含有1个残基/分子的tritriated hypusine （N - epsilon-（4-氨基-2-羟基丁基）赖氨酸），这是一种罕见的氨基酸，在其他蛋白质中没有发现。为了研究这种蛋白的保守性，我们检测了两种非哺乳动物真核生物，酵母酿酒酵母和昆虫黑腹果蝇，以及真细菌原核生物大肠杆菌中含有这种含hypusine蛋白的存在。当真核细胞在[3H]亚精胺存在下生长时，电泳分析显示单个标记蛋白。每一种蛋白的表观分子量均接近18000，相对pI约为5.2，与哺乳动物的含hypusine蛋白相似。氨基酸分析证实，在每种情况下都存在氚化的hypusine，二维聚丙烯酰胺凝胶的银染色表明，酵母和果蝇与哺乳动物一样，这种蛋白质相对丰富。在大肠杆菌中，一种氚化蛋白占主导地位，但其分子量为24,000，我们没有发现它含有氚化hypusine的证据。我们没有发现在古细菌伏曲甲烷球菌中存在含hypusine蛋白的证据。这些数据表明含有hypusine的蛋白在真核生物中是保守的。
When mammalian cells are grown in medium containing [3H]spermidine, a single major tritiated protein identical to eukaryotic initiation factor 4D becomes labeled. This protein contains 1 residue/molecule of tritiated hypusine (N epsilon-(4-amino-2-hydroxybutyl)lysine), a rare amino acid which has been found in no other protein. In order to investigate the conservation of this protein, we examined two nonmammalian eukaryotes, the yeast Saccharomyces cerevisiae and the insect Drosophila melanogaster, and the eubacterial prokaryote Escherichia coli for the presence of the hypusine-containing protein. When the eukaryotic cells were grown in the presence of [3H]spermidine, electrophoretic analysis revealed a single labeled protein. In each case, the apparent molecular weight was near 18,000 and the relative pI was approximately 5.2, similar to the hypusine-containing protein of mammals. Amino acid analysis confirmed the presence of tritiated hypusine in each case, and silver staining of two-dimensional polyacrylamide gels demonstrated that, in yeast and fruit flies as in mammals, the protein is relatively abundant. In the eubacterium E. coli, one tritiated protein was predominant, but its molecular weight was 24,000 and we found no evidence that it contained tritiated hypusine. We found no evidence for the existence of the hypusine-containing protein in the archaebacterium Methanococcus voltae. These data suggest that the hypusine-containing protein is conserved among eukaryotes.