Tateishi-Karimata Hisae, Nakano Shu-Ichi, Sugimoto Naoki
Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University, Kobe, Japan.
Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, Kobe, Japan.
Curr Protoc Nucleic Acid Chem. 2013 Jun;Chapter 7:7.19.1-7.19.17. doi: 10.1002/0471142700.nc0719s53.
A variety of biomolecules, including nucleic acids, proteins, polysaccharides, and other soluble and insoluble low-molecular weight components, are present in living cells. These molecules occupy a significant fraction of the cellular volume (up to 40%), resulting in a highly crowded intracellular environment. This situation is referred to as molecular crowding. Although the thermodynamic stabilities of DNA structures are known to be altered in a crowded environment, less is known about the behavior of nucleic acids and their interactions with cations and water molecules under such conditions. This unit describes methods that can be used to quantitatively analyze the molecular crowding effects caused by cosolutes on the thermodynamic stability, hydration, and cation binding of nucleic acid structures.
活细胞中存在多种生物分子，包括核酸、蛋白质、多糖以及其他可溶性和不可溶性的低分子量成分。这些分子占据了细胞体积的很大一部分（高达40%），导致细胞内环境高度拥挤。这种情况被称为分子拥挤。尽管已知在拥挤环境中DNA结构的热力学稳定性会发生改变，但在这种条件下核酸的行为及其与阳离子和水分子的相互作用却知之甚少。本单元介绍了可用于定量分析共溶质对核酸结构的热力学稳定性、水合作用和阳离子结合所造成的分子拥挤效应的方法。
