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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Weak protein-cationic co-ion interactions addressed by X-ray crystallography and mass spectrometry
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Weak protein-cationic co-ion interactions addressed by X-ray crystallography and mass spectrometry

机译：弱protein-cationic co-ion交互通过x射线晶体学和质量光谱法

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摘要

The adsorption of Rb+, Cs+, Mn2+, Co2+ and Yb3+ onto the positively charged hen egg-white lysozyme (HEWL) has been investigated by solving 13 X-ray structures of HEWL crystallized with their chlorides and by applying electrospray ionization mass spectrometry (ESI-MS) first to dissolved protein crystals and then to the protein in buffered salt solutions. The number of bound cations follows the order Cs+ < Mn2+ similar or equal to Co2+ < Yb3+ at 293 K. HEWL binds less Rb+ (q(tot) = 0.7) than Cs+ (q(tot) = 3.9) at 100 K. Crystal flash-cooling drastically increases the binding of Cs+, but poorly affects that of Yb3+, suggesting different interactions. The addition of glycerol increases the number of bound Yb3+ cations, but only slightly increases that of Rb+. HEWL titrations with the same chlorides, followed by ESI-MS analysis, show that only about 10% of HEWL binds Cs+ and about 40% binds 1-2 Yb3+ cations, while the highest binding reaches 60-70% for protein binding 1-3 Mn2+ or Co2+ cations. The binding sites identified by X-ray crystallography show that the monovalent Rb+ and Cs+ preferentially bind to carbonyl groups, whereas the multivalent Mn2+, Co2+ and Yb3+ interact with carboxylic groups. This work elucidates the basis of the effect of the Hofmeister cation series on protein solubility.
机译：《Rb adsorption Cs +、Mn2 +、Co2和Yb3 +到母鸡带正电蛋白溶菌酶(HEWL)已经被解决了13 x射线结构HEWL结晶氯化物和通过应用电喷射电喷雾质谱电离质谱()第一溶解的蛋白质晶体,然后蛋白质在缓冲盐的解决方案。结合Cs + < Mn2 +阳离子遵循秩序类似或等于Co2 + < Yb3 + 293 K。结合少Rb + (q(合计)= 0.7)比c + (q(合计)=3.9)在100 K。增加了c +的绑定,但不影响Yb3 +,表明不同的交互。甘油的加入增加的数量绑定Yb3 +阳离子,但仅略有增加Rb +。氯化物,紧随其后的是谱分析,证明只有约10%的HEWL结合Cs +和40%左右结合1 - 2 Yb3 +阳离子,而最高的绑定达到60 - 70%蛋白质绑定1 - 3 Mn2 +或二氧化碳+阳离子。x射线晶体学表明,单价铷+和+优先绑定到羰基组,而多价Mn2 + +和二氧化碳Yb3 +与羧基组。拇外翻的效果的基础他认为系列阳离子蛋白质溶解度。

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