Laboratory of Biophysics, Solid State Physics Department, Jozef Stefan Institute, 1000 Ljubljana, Slovenia.
Eur Biophys J. 2010 Mar;39(4):499-511. doi: 10.1007/s00249-009-0510-5. Epub 2009 Aug 11.
As proteins are key molecules in living cells, knowledge about their structure can provide important insights and applications in science, biotechnology, and medicine. However, many protein structures are still a big challenge for existing high-resolution structure-determination methods, as can be seen in the number of protein structures published in the Protein Data Bank. This is especially the case for less-ordered, more hydrophobic and more flexible protein systems. The lack of efficient methods for structure determination calls for urgent development of a new class of biophysical techniques. This work attempts to address this problem with a novel combination of site-directed spin labelling electron spin resonance spectroscopy (SDSL-ESR) and protein structure modelling, which is coupled by restriction of the conformational spaces of the amino acid side chains. Comparison of the application to four different protein systems enables us to generalize the new method and to establish a general procedure for determination of protein structure.
由于蛋白质是活细胞中的关键分子，因此了解其结构可以为科学、生物技术和医学提供重要的见解和应用。然而，许多蛋白质结构仍然是现有高分辨率结构测定方法的一大挑战，这可以从蛋白质数据库中发表的蛋白质结构数量看出。对于较无序、疏水性更强和更灵活的蛋白质系统尤其如此。缺乏有效的结构测定方法要求迫切需要开发一类新的生物物理技术。这项工作试图通过一种新颖的组合来解决这个问题，即定点旋转标记电子自旋共振光谱学（SDSL-ESR）和蛋白质结构建模，该组合通过限制氨基酸侧链的构象空间来实现。将该方法应用于四个不同的蛋白质系统的比较使我们能够推广新方法，并建立确定蛋白质结构的一般程序。
