Different mechanisms of action of poly(ethylene glycol) and arginine on thermal inactivation of lysozyme and ribonuclease A
Different mechanisms of action of poly(ethylene glycol) and arginine on thermal inactivation of lysozyme and ribonuclease A
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聚乙二醇和精氨酸对溶菌酶和核糖核酸酶 A 热失活的不同作用机制
DOI：
10.1002/bit.24531
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发表时间：
2012
影响因子：
3.8
通讯作者：
Shunsuke Tomita
中科院分区：
文献类型：
--
作者：
Otsuka T;Kawaguchi Y;末水洋志;Shunsuke Tomita
蛋白质往往通过几种化学修饰而发生不可逆的失活，这在各个领域都是一个严重的问题。我们最近发现，精氨酸（Arg）抑制热诱导的脱酰胺和β消除，从而抑制鸡蛋白色溶菌酶和牛胰腺核糖核酸酶A的热灭活。在这里，我们报告说，聚（乙二醇）（PEG）与分子量为1,000作为两种蛋白质的热失活抑制剂，特别是在较高的蛋白质浓度，而精氨酸在较高的蛋白质浓度是无效的。这种差异表明，PEG，而不是精氨酸，有效地抑制热变性蛋白质之间的分子间二硫键交换。研究了各种聚合物，包括不同分子量的PEG、聚（乙烯基吡咯烷酮）（PVP）和聚（乙烯醇）对蛋白质热失活的影响、圆二色性、溶液粘度以及还原和S-羧甲基化溶菌酶的溶解度表明，两亲性PEG和PVP通过优先与热变性蛋白质相互作用来抑制热变性蛋白质的分子间碰撞，导致分子间二硫键交换的抑制。这些关于两亲性聚合物-PEG和PVP-和Arg的不同作用机制的发现将扩大提高溶液中蛋白质化学稳定性的方法的能力。Biotechnol. Bioeng. 2012; 109：2543-2552.© 2012 Wiley Periodicals，Inc.
Proteins tend to undergo irreversible inactivation through several chemical modifications, which is a serious problem in various fields. We have recently found that arginine (Arg) suppresses heat‐induced deamidation and β‐elimination, resulting in the suppression of thermal inactivation of hen egg white lysozyme and bovine pancreas ribonuclease A. Here, we report that poly(ethylene glycol) (PEG) with molecular weight 1,000 acts as a thermoinactivation suppressor for both proteins, especially at higher protein concentrations, while Arg was not effective at higher protein concentrations. This difference suggests that PEG, but not Arg, effectively inhibited intermolecular disulfide exchange among thermally denatured proteins. Investigation of the effects of various polymers including PEG with different molecular weight, poly(vinylpyrolidone) (PVP), and poly(vinyl alchol) on thermoinactivation of proteins, circular dichroism, solution viscosity, and the solubility of reduced andS‐carboxy‐methylated lysozyme indicated that amphiphilic PEG and PVP inhibit intermolecular collision of thermally denatured proteins by preferential interaction with thermally denatured proteins, resulting in the inhibition of intermolecular disulfide exchange. These findings regarding the different mechanisms of the effects of amphiphilic polymers––PEG and PVP––and Arg would expand the capabilities of methods to improve the chemical stability of proteins in solution. Biotechnol. Bioeng. 2012; 109: 2543–2552. © 2012 Wiley Periodicals, Inc.