Ingrosso D, Fowler A V, Bleibaum J, Clarke S
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1569.
J Biol Chem. 1989 Nov 25;264(33):20131-9.
A widely distributed protein methyltransferase catalyzes the transfer of a methyl group from S-adenosyl-methionine to the free carboxyl groups of D-aspartyl and/or L-isoaspartyl derivatives of L-aspartyl and L-asparaginyl residues. This enzyme has been postulated to function in the repair or the catabolism of age-damaged proteins. We present here the complete amino acid sequence of the more basic isozyme I of this enzyme from human erythrocytes. The sequence was determined by Edman degradation and mass spectral analysis of overlapping trypsin, Staphylococcus aureus V8 protease, Pseudomonas fragi endoproteinase Asp-N, cyanogen bromide, and hydroxylamine-generated fragments. The NH2-terminus is modified by acetylation and the protein contains 226 amino acids for a calculated molecular weight of 24,575. This value is in good agreement with the molecular weight determined for the purified protein by polyacrylamide gel electrophoresis in the presence of dodecyl sulfate and by gel filtration chromatography under nondenaturing conditions. The identification of 2 different amino acid residues at both positions 22 and 119 may indicate the presence of allelic variants or of two or more closely related structural genes. Finally, comparison of this sequence with those of methyltransferases for RNA, DNA, and small molecules, as well as other S-adenosylmethionine-utilizing enzymes, shows that many of these proteins share elements of three regions of sequence similarity and may be structurally or evolutionarily related.
一种广泛分布的蛋白质甲基转移酶催化甲基从S-腺苷甲硫氨酸转移至L-天冬氨酰和L-天冬酰胺残基的D-天冬氨酰和/或L-异天冬氨酰衍生物的游离羧基上。据推测,这种酶在衰老受损蛋白质的修复或分解代谢中发挥作用。我们在此展示了来自人红细胞的该酶碱性更强的同工酶I的完整氨基酸序列。该序列通过对胰蛋白酶、金黄色葡萄球菌V8蛋白酶、脆弱拟杆菌天冬氨酸内肽酶Asp-N、溴化氰和羟胺产生的重叠片段进行埃德曼降解和质谱分析来确定。氨基末端被乙酰化修饰,该蛋白质包含226个氨基酸,计算分子量为24,575。这个值与在十二烷基硫酸钠存在下通过聚丙烯酰胺凝胶电泳以及在非变性条件下通过凝胶过滤色谱法测定的纯化蛋白质的分子量高度一致。在第22位和第119位均鉴定出2种不同的氨基酸残基,这可能表明存在等位基因变体或两个或更多密切相关的结构基因。最后,将该序列与RNA、DNA和小分子的甲基转移酶以及其他利用S-腺苷甲硫氨酸的酶的序列进行比较,结果表明这些蛋白质中有许多共享三个序列相似区域的元件,可能在结构上或进化上相关。