Adenylate kinase from Sulfolobus acidocaldarius: Expression in Escherichia coli and characterization by Fourier transform infrared spectroscopy
Adenylate kinase from Sulfolobus acidocaldarius: Expression in Escherichia coli and characterization by Fourier transform infrared spectroscopy
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DOI：
10.1006/abbi.1996.0366
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发表时间：
1996-09-01
影响因子：
3.9
通讯作者：
Schafer, G
中科院分区：
文献类型：
--
作者：
Bonisch, H;Backmann, J;Schafer, G
从极度嗜热嗜酸的古细菌Sulfolobus acidocalarius中提取的腺苷酸激酶在大肠杆菌中过度表达。用傅里叶变换红外光谱（FTIR）对高纯度酶进行了表征。FTIR光谱分析和二级结构估计表明，该蛋白的整体结构与其他腺苷酸激酶相似。由于蛋白质聚集，蛋白质的热展开（估计T-m值接近90℃）是不可逆的。该酶在80℃下表现出长期稳定性，对75-80℃的生理生长温度具有良好的适应性。二级结构敏感带的半宽度和氢-氘交换实验表明，与猪肌细胞质中的腺苷酸激酶相比，Sulfolobus酶具有明显更紧凑和刚性的蛋白核心结构。这可能特别有助于蛋白质的极端热稳定性。(C) 1996学术出版社，Inc.
Adenylate kinase from the extremely thermoacidophilic archaeon Sulfolobus acidocaldarius has been overexpressed in Escherichia coli. The highly purified enzyme was characterized by Fourier transform infrared spectroscopy (FTIR). Analysis of FTIR spectra and estimation of secondary structure revealed a global protein structure similar to that of other adenylate kinases. Thermal unfolding of the protein with an estimated T-m value near 90 degrees C is irreversible due to protein aggregation. The enzyme exhibits long-term stability up to 80 degrees C, which is an excellent adaptation to the physiological growth temperature of 75-80 degrees C. Halfwidths of secondary-structure-sensitive bands and hydrogen-deuterium exchange experiments revealed that in comparison to adenylate kinase from porcine muscle cytosol the Sulfolobus enzyme is characterized by a significantly more compact and rigid protein core structure, which is likely to contribute specifically to the extreme thermostability of the protein. (C) 1996 Academic Press, Inc.