Identification of yeast aspartyl aminopeptidase gene by purifying and characterizing its product from yeast cells
Identification of yeast aspartyl aminopeptidase gene by purifying and characterizing its product from yeast cells
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DOI：
10.1111/j.1742-4658.2005.05057.x
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发表时间：
2006-01
期刊：
影响因子：
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通讯作者：
Ryo Yokoyama;H. Kawasaki;H. Hirano
中科院分区：
文献类型：
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作者：
Ryo Yokoyama;H. Kawasaki;H. Hirano
天冬氨酰氨基肽酶（EC 3.4.11.21）仅切割未封闭的N末端酸性氨基酸残基。迄今为止，它仅在哺乳动物中发现。我们在这里报告，乙酰氨基肽酶活性是存在于酵母。酵母氨肽酶由染色体VIII中的未表征基因（YHR 113 W，酵母基因组数据库）编码。酵母乙酰氨基肽酶优先切割肽的未封闭的N末端酸性氨基酸残基;该活性的最佳pH在中性范围内。金属蛋白酶抑制剂EDTA和1.10-菲咯啉均抑制酶的活性，而大多数氨肽酶的抑制剂bestatin不影响酶活性。凝胶过滤色谱法显示，酵母乙酰氨基肽酶的天然形式的分子量为1680 000。纯化的酵母乙酰氨基肽酶的SDS/PAGE产生单个56-kDa条带，表明该酶包含12个相同的亚基。
Aspartyl aminopeptidase (EC 3.4.11.21) cleaves only unblocked N‐terminal acidic amino‐acid residues. To date, it has been found only in mammals. We report here that aspartyl aminopeptidase activity is present in yeast. Yeast aminopeptidase is encoded by an uncharacterized gene in chromosome VIII (YHR113W, Saccharomyces Genome Database). Yeast aspartyl aminopeptidase preferentially cleaved the unblocked N‐terminal acidic amino‐acid residue of peptides; the optimum pH for this activity was within the neutral range. The metalloproteases inhibitors EDTA and 1.10‐phenanthroline both inhibited the activity of the enzyme, whereas bestatin, an inhibitor of most aminopeptidases, did not affect enzyme activity. Gel filtration chromatography revealed that the molecular mass of the native form of yeast aspartyl aminopeptidase is ≈ 680 000. SDS/PAGE of purified yeast aspartyl aminopeptidase produced a single 56‐kDa band, indicating that this enzyme comprises 12 identical subunits.