Kornblatt Jack A, Barretto Tanya A, Chigogidze Ketevan, Chirwa Bahati
Enzyme Research Group, Department of Biology, Concordia University, Montreal, Qc., Canada.
Anal Chem Insights. 2007 Mar 22;2:17-29.
We studied the near UV absorption spectrum of canine plasminogen. There are 19 tryptophans, 19 phenylalanines and 34 tyrosines in the protein. 4th derivative spectra optimized for either tryptophan or tyrosine give a measure of the polarity of the environments of these two aromatic amino acids. Plasminogen at temperatures between 0 degrees C and 37 degrees C exists as a mixture of four conformations: closed-relaxed, open-relaxed, closed-compact, and open-compact. The closed to open transition is driven by addition of ligand to a site on the protein. The relaxed to compact transition is driven by increasing temperature from 0 degrees C to above 15-20 degrees C.When the conformation of plasminogen is mainly closed-relaxed, the 4th derivative spectra suggest that the average tryptophan environment is similar to a solution of 20% methanol at the same temperature. Under the same conditions, 4th derivative spectra suggest that the average tyrosine environment is similar to water. These apparent polarities change as the plasminogen is forced to assume the other conformations. We try to rationalize the information based on the known portions of the plasminogen structure.
我们研究了犬纤溶酶原的近紫外吸收光谱。该蛋白质中有19个色氨酸、19个苯丙氨酸和34个酪氨酸。针对色氨酸或酪氨酸优化的四阶导数光谱可衡量这两种芳香族氨基酸所处环境的极性。在0摄氏度至37摄氏度之间的温度下,纤溶酶原以四种构象的混合物形式存在:闭合-松弛型、开放-松弛型、闭合-紧密型和开放-紧密型。从闭合到开放的转变是由向蛋白质上的一个位点添加配体驱动的。从松弛到紧密的转变是由温度从0摄氏度升高到15-20摄氏度以上驱动的。当纤溶酶原的构象主要为闭合-松弛型时,四阶导数光谱表明,平均色氨酸环境与相同温度下20%甲醇溶液相似。在相同条件下,四阶导数光谱表明,平均酪氨酸环境与水相似。当纤溶酶原被迫呈现其他构象时,这些表观极性会发生变化。我们试图根据纤溶酶原结构的已知部分对这些信息进行合理分析。