Biochemical properties and primary structure of elastase inhibitor AFUEI from Aspergillus fumigatus.
Biochemical properties and primary structure of elastase inhibitor AFUEI from Aspergillus fumigatus.
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烟曲霉弹性蛋白酶抑制剂 AFUEI 的生化特性和一级结构。
DOI：
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发表时间：
2008
影响因子：
3
通讯作者：
T. Nikai
中科院分区：
文献类型：
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作者：
Y. Okumura;Takeshi Matsui;K. Ogawa;K. Uchiya;T. Nikai
从烟曲霉中分离得到一种弹性蛋白酶抑制剂AFUEI，并对其生化性质和一级结构进行了研究。用DE 52纤维素和Sephadex G-75柱层析纯化抑制剂，发现其为均一的，如不连续PAGE和SDS-PAGE后的单一条带所示。通过基质辅助解吸/电离飞行时间质谱法观察到7525.1 Da的分子量。对A. AFUEI抑制烟曲霉、黄曲霉和人白细胞的增殖。然而，猪胰腺弹性蛋白酶、绿脓杆菌弹性蛋白酶和蛇毒弹性蛋白酶的弹性蛋白溶解活性不受AFUEI的影响。DTT或2-巯基乙醇对AFUEI的弹性蛋白酶抑制活性无抑制作用。通过Edman测序确定源自利用梭菌蛋白酶的细菌的AFUEI肽的氨基酸序列。AFUEI由68个氨基酸组成，计算分子量为7526.2Da，氨基酸同源性分析表明AFUEI的氨基酸1 - 68与A.烟熏。
An elastase inhibitor from Aspergillus fumigatus (AFUEI) was isolated, and its biochemical properties and primary structure examined. The inhibitor was purified by column chromatography using DE52 cellulose and Sephadex G-75, and was found to be homogeneous as indicated by a single band following discontinuous PAGE and SDS-PAGE. A molecular mass of 7525.1 Da was observed by matrix-assisted desorption/ionization time-of-flight mass spectroscopy. The elastolytic activity of elastases from A. fumigatus, Aspergillus flavus and human leukocytes was inhibited by AFUEI. However, the elastolytic activity of porcine pancreas elastase, Pseudomonas aeruginosa elastase and elastase from snake venom was not affected by AFUEI. No inhibitory effect of DTT or 2-mercaptoethanol on the elastase inhibitory activity of AFUEI was observed. The amino acid sequence of AFUEI peptides derived from digests utilizing clostripain was determined by Edman sequencing. AFUEI was composed of 68 aa and had a calculated molecular mass of 7526.2 Da. The search for amino acid homology with other proteins demonstrated that aa 1-68 of AFUEI are 100 % identical to aa 20-87 of the hypothetical protein AFUA 3G14940 of A. fumigatus.