Interdepartmental Program in Biomolecular Science and Engineering, University of California, Santa Barbara, California, 93106.
Protein Sci. 2014 Jan;23(1):56-66. doi: 10.1002/pro.2390. Epub 2013 Nov 28.
The folding of larger proteins generally differs from the folding of similarly large nucleic acids in the number and stability of the intermediates involved. To date, however, no similar comparison has been made between the folding of smaller proteins, which typically fold without well-populated intermediates, and the folding of small, simple nucleic acids. In response, in this study, we compare the folding of a 38-base DNA aptamer with the folding of a set of equivalently simple proteins. We find that, as is true for the large majority of simple, single domain proteins, the aptamer folds through a concerted, millisecond-scale process lacking well-populated intermediates. Perhaps surprisingly, the observed folding rate falls within error of a previously described relationship between the folding kinetics of single-domain proteins and their native state topology. Likewise, similarly to single-domain proteins, the aptamer exhibits a relatively low urea-derived Tanford β, suggesting that its folding transition state is modestly ordered. In contrast to this, however, and in contrast to the behavior of proteins, ϕ-value analysis suggests that the aptamer's folding transition state is highly ordered, a discrepancy that presumably reflects the markedly more important role that secondary structure formation plays in the folding of nucleic acids. This difference notwithstanding, the similarities that we observe between the two-state folding of single-domain proteins and the two-state folding of this similarly simple DNA presumably reflect properties that are universal in the folding of all sufficiently cooperative heteropolymers irrespective of their chemical details.
较大蛋白质的折叠通常与类似大小的核酸的折叠不同,涉及的中间体的数量和稳定性不同。然而,迄今为止,在较小蛋白质的折叠(通常没有形成良好的中间体)与较小简单核酸的折叠之间,尚未进行类似的比较。有鉴于此,在这项研究中,我们比较了一个 38 个碱基的 DNA 适体与一组同样简单的蛋白质的折叠。我们发现,与绝大多数简单的单一结构域蛋白质一样,适体通过协同的毫秒级过程折叠,没有形成良好的中间体。也许令人惊讶的是,观察到的折叠速率与以前描述的单结构域蛋白质的折叠动力学与其天然状态拓扑结构之间的关系误差范围内。同样,与单结构域蛋白质相似,适体表现出相对较低的尿素衍生的 Tanford β,表明其折叠转变态适度有序。然而,与这一点形成对比的是,与蛋白质的行为相反,ϕ 值分析表明适体的折叠转变态高度有序,这种差异可能反映了二级结构形成在核酸折叠中起着更为重要的作用。尽管存在这种差异,但我们观察到单结构域蛋白质的两态折叠与这种同样简单的 DNA 的两态折叠之间的相似之处,可能反映了所有足够协同的杂聚物在折叠中普遍存在的特性,而不论其化学细节如何。