通过荧光猝灭揭示的蛋白质基质动力学
Dynamics of a protein matrix revealed by fluorescence quenching.
作者信息
Eftink M R, Ghiron C A
出版信息
Proc Natl Acad Sci U S A. 1975 Sep;72(9):3290-4. doi: 10.1073/pnas.72.9.3290.
Abstract
The fluorescence of the supposedly buried tryptophan in ribonuclease T1 has been found to be collisionally quenched by acrylamide with a rate constant of 3 X 10(8) M--1 sec--1. Only a slight decrease in the quenching rate is observed upon a 5-fold increase in the viscosity of the solution. For this to be the case, the diffusion of the quencher must be limited by the protein matrix. To explain the process of diffusion through this complex material, the formation of "holes" in the lattice of a protein due to nanosecond fluctuations must be invoked. Thus, the dynamic character of a protein molecule is revealed. The quenching rate constant has an activation energy of 9 kcal/mol which can be used to characterize the nature of the cohesive forces in the microenvironment about the indole ring. The mechanical properties of a portion of a protein matrix can, therefore, be described as one would for a fluid.
摘要
已发现核糖核酸酶T1中假定被掩埋的色氨酸的荧光会被丙烯酰胺碰撞猝灭，猝灭速率常数为3×10⁸ M⁻¹ s⁻¹。溶液粘度增加5倍时，仅观察到猝灭速率略有下降。要出现这种情况，猝灭剂的扩散必定受到蛋白质基质的限制。为了解释通过这种复杂物质的扩散过程，必须借助由于纳秒级波动而在蛋白质晶格中形成的“空洞”。因此，揭示了蛋白质分子的动态特性。猝灭速率常数的活化能为9千卡/摩尔，这可用于表征吲哚环周围微环境中内聚力的性质。因此，蛋白质基质一部分的机械性能可以像描述流体那样来描述。
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本文引用的文献
1
The structure and function of ribonuclease T1. II. Further purification and amino acid composition of ribonuclease T1.核糖核酸酶T1的结构与功能。II. 核糖核酸酶T1的进一步纯化及氨基酸组成
J Biochem. 1962 Feb;51:95-108. doi: 10.1093/oxfordjournals.jbchem.a127515.
2
Further observations on the electrical properties of hemoglobin-bound water.关于血红蛋白结合水的电学性质的进一步观察
J Phys Chem. 1969 Aug;73(8):2600-10. doi: 10.1021/j100842a024.
3
The luminescence of tryptophan and phenylalanine derivatives.色氨酸和苯丙氨酸衍生物的发光
Biochemistry. 1968 Jul;7(7):2488-95. doi: 10.1021/bi00847a007.
4
Conformation and segmental motion of native and denatured ribonuclease A in solution. Application of natural-abundance carbon-13 partially relaxed Fourier transform nuclear magnetic resonance.溶液中天然和变性核糖核酸酶A的构象与片段运动。天然丰度碳-13部分弛豫傅里叶变换核磁共振的应用。
J Am Chem Soc. 1971 Jan 27;93(2):544-6. doi: 10.1021/ja00731a053.
5
6
On the thermodynamic basis of induced fit. Specific alkane binding to proteins.基于诱导契合的热力学原理。特定烷烃与蛋白质的结合。
Biochemistry. 1969 Dec;8(12):5070-5. doi: 10.1021/bi00840a059.
7
Solute perturbation of protein fluorescence. The quenching of the tryptophyl fluorescence of model compounds and of lysozyme by iodide ion.蛋白质荧光的溶质扰动。碘离子对模型化合物和溶菌酶色氨酸荧光的猝灭作用。
Biochemistry. 1971 Aug 17;10(17):3254-63. doi: 10.1021/bi00793a015.
8
Nanosecond time-resolved fluorescence spectra of a protein-dye complex.蛋白质 - 染料复合物的纳秒时间分辨荧光光谱。
J Biol Chem. 1971 Apr 10;246(7):2317-9.
9
Enthalpy-entropy compensation phenomena in water solutions of proteins and small molecules: a ubiquitous property of water.蛋白质和小分子水溶液中的焓-熵补偿现象：水的一种普遍特性。
Biopolymers. 1970;9(10):1125-227. doi: 10.1002/bip.1970.360091002.
10
Resonance energy transfer in pepsin conjugates.胃蛋白酶共轭物中的共振能量转移。
J Mol Biol. 1969 Aug 28;44(1):71-88. doi: 10.1016/0022-2836(69)90405-7.
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