Ivanov Yuri D, Govorun Vadim M, Bykov Victor A, Archakov Alexander I
Institute of Biomedical Chemistry, Moscow, Russia.
Proteomics. 2006 Mar;6(5):1399-414. doi: 10.1002/pmic.200402087.
Progress in proteomic researches is largely determined by development and implementation of new methods for the revelation and identification of proteins in biological material in a wide concentration range (from 10(-3) M to single molecules). The most perspective approaches to address this problem involve (i) nanotechnological physicochemical procedures for the separation of multicomponent protein mixtures; among these of particular interest are biospecific nanotechnological procedures for selection of proteins from multicomponent protein mixtures with their subsequent concentration on solid support; (ii) identification and counting of single molecules by use of molecular detectors. The prototypes of biospecific nanotechnological procedures, based on the capture of ligand biomolecules by biomolecules of immobilized ligate and the concentration of the captured ligands on appropriate surfaces, are well known; these are affinity chromatography, magnetic biobeads technology, different biosensor methods, etc. Here, we review the most promising nanotechnological approaches for selection of proteins and kinetic characterization of their complexes based on these biospecific methods with subsequent MS/MS identification of proteins and protein complexes. Two major groups of methods for the analysis and identification of individual molecules and their complexes by use of molecular detectors will be reviewed: scanning probe microscopy (SPM) (including atomic-force microscopy) and cryomassdetector technology.
蛋白质组学研究的进展在很大程度上取决于新方法的开发与应用,这些新方法用于揭示和鉴定生物材料中浓度范围广泛(从10⁻³ M到单分子)的蛋白质。解决这一问题最具前景的方法包括:(i)用于分离多组分蛋白质混合物的纳米技术物理化学程序;其中特别令人感兴趣的是生物特异性纳米技术程序,用于从多组分蛋白质混合物中选择蛋白质,并随后将其浓缩在固体支持物上;(ii)使用分子探测器对单分子进行鉴定和计数。基于固定化连接子的生物分子捕获配体生物分子并将捕获的配体浓缩在合适表面上的生物特异性纳米技术程序的原型是众所周知的;这些方法包括亲和色谱法、磁性生物珠技术、不同的生物传感器方法等。在此,我们综述了基于这些生物特异性方法选择蛋白质及其复合物的动力学表征,随后通过MS/MS鉴定蛋白质和蛋白质复合物的最有前景的纳米技术方法。将综述利用分子探测器分析和鉴定单个分子及其复合物的两大类方法:扫描探针显微镜(SPM)(包括原子力显微镜)和低温质量探测器技术。