Alliance Protein Laboratories, 3957 Corte Cancion, Thousand Oaks, CA 91360, USA.
Int J Biol Macromol. 2010 Mar 1;46(2):217-22. doi: 10.1016/j.ijbiomac.2009.11.007. Epub 2009 Dec 3.
Aqueous arginine solution now finds a wide range of applications in biotechnology fields, including protein refolding, chromatography and virus inactivation. While progress has been made for mechanistic understanding of the effects of arginine on proteins, we have little understanding on how arginine inactivates viruses. One of the viral components is nucleic acid. We have examined the effects of arginine on the structure and thermal stability of calf thymus deoxyribonucleic acid (DNA) using circular dichroism (CD). Both NaCl and arginine reduced CD intensity. At low concentrations, arginine showed a stronger effect on CD intensity than NaCl. Both NaCl and arginine sharply increased the melting temperature at low concentrations (below 0.25 M). However, they had an opposite effect at higher concentrations. Above this concentration, NaCl gradually increased the melting temperature, leading to the onset melting temperature above 90 degrees C. On the other hand, the thermal stability in the presence of arginine reached a maximum at 0.2-0.5 M, after which further addition of arginine caused decreased melting temperature. It is most likely that the increased melting temperature at low concentration is due to electrostatic stabilization of DNA structure by both NaCl and arginine and that the opposite effects at higher salt concentration are due to salt-specific effects, i.e., stabilizing (salting-out) effects of NaCl and destabilizing (salting-in) effects of arginine. Solubility measurements of nucleic acid bases showed that arginine, but not NaCl, increases the solubilities of the bases, supporting their effects on DNA stability at higher concentration.
氨基酸溶液在生物技术领域有广泛的应用,包括蛋白质复性、色谱和病毒失活。虽然人们对精氨酸影响蛋白质的机制有了一定的了解,但对精氨酸如何使病毒失活知之甚少。病毒的一个组成部分是核酸。我们用圆二色性(CD)法研究了精氨酸对小牛胸腺脱氧核糖核酸(DNA)结构和热稳定性的影响。NaCl 和精氨酸都降低了 CD 强度。在低浓度下,精氨酸对 CD 强度的影响比 NaCl 更强。NaCl 和精氨酸都在低浓度(低于 0.25M)急剧增加了熔点。然而,在较高浓度下,它们的效果相反。在这个浓度以上,NaCl 逐渐增加了熔点,导致起始熔点高于 90 摄氏度。另一方面,在精氨酸存在下,热稳定性在 0.2-0.5M 达到最大值,之后再加入精氨酸会导致熔点降低。很可能低浓度下熔点升高是由于 NaCl 和精氨酸静电稳定了 DNA 结构,而在较高盐浓度下的相反效果是由于盐的特异性效应,即 NaCl 的稳定(盐析)效应和精氨酸的不稳定(盐溶)效应。核酸碱基的溶解度测量表明,精氨酸而非 NaCl 增加了碱基的溶解度,这支持了它们在较高浓度下对 DNA 稳定性的影响。