DETERMINATION OF PROTEIN SECONDARY STRUCTURE BY FOURIER-TRANSFORM INFRARED-SPECTROSCOPY - A CRITICAL-ASSESSMENT
DETERMINATION OF PROTEIN SECONDARY STRUCTURE BY FOURIER-TRANSFORM INFRARED-SPECTROSCOPY - A CRITICAL-ASSESSMENT
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DOI：
10.1021/bi00053a001
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发表时间：
1993-01-19
期刊：
影响因子：
2.9
通讯作者：
CHAPMAN, D
中科院分区：
文献类型：
--
作者：
SUREWICZ, WK;MANTSCH, HH;CHAPMAN, D
蛋白质结构研究的最终目的是在高分辨率水平上洞察蛋白质的三维结构。这通常可以通过应用诸如X射线结晶学或多维核磁共振(核磁共振)光谱等技术来实现。然而，对蛋白质的高分辨率研究并不总是可行的。例如，结晶学研究需要高质量的单晶，而对于许多蛋白质(例如，绝大多数膜蛋白)来说，这是不存在的。此外，单晶体中相对“静态”的结构是否充分代表了活细胞复杂而动态的环境中的蛋白质构象，这也是一个问题。越来越多的人认识到[例如，Martinek等人()]在体内，大多数蛋白质在界面环境中发挥作用，在那里它们与生物膜、核酸、多糖或其他蛋白质形成动态复合体。通常用来生长蛋白质晶体的水溶液不一定能很好地模拟蛋白质在体内发挥作用的条件。核磁共振在研究“生物相关”环境中的蛋白质结构方面提供了更好的灵活性。然而，较大蛋白质的核磁共振谱的解释非常复杂，由核磁共振实验产生的质子间距离的分配并不总是可行的；目前该技术仅限于小于约15-20 kDa的小蛋白质。在蛋白质的高分辨率结构研究中遇到的实际限制刺激了在这方面的持续进展。NRCC
The ultimate goal of structural studies of proteins is to gain insight into protein three-dimensional structure at highresolution level. This can often be accomplished by the application of techniques such as X-ray crystallography or multidimensional nuclear magnetic resonance (NMR) spectroscopy. However, high-resolution studies of proteins are not always feasible. For example, crystallographic studies require high-quality single crystals which for many proteins (eg, the vast majority of membrane proteins) are not available. Furthermore, the question arises as to whether the relatively “static” structure in single crystals adequately represents the protein conformation in a complex and dynamic environment of living cells. There isa growing realization [eg, Martinek et al.(1989)] that invivo most proteins act in an interfacial environment where they form dynamic complexes with biological membranes, nucleic acids, polysaccharides, or other proteins. Aqueous buffers, from which protein crystals are usually grown, do not necessarily mimic well the conditions of protein functioning in vivo. NMR offers a somewhat better flexibility in studying protein structure in “biologically relevant” environments. However, the interpretation ofNMR spectra of larger proteins is very complex, and the assignment of interproton distances generated by the NMR experiment is not always feasible; at present the technique is restricted to small proteins of less than approximately 15-20 kDa. The practical limitations encountered in high-resolution structural studies of proteins stimulate continual progress in f This is National Research Councilof Canada PublicationNo. NRCC