Grell D, Richardson J S, Mutter M
Institute of Organic Chemistry, University of Lausanne, Switzerland.
J Pept Sci. 2001 Mar;7(3):146-51. doi: 10.1002/psc.308.
An experimental system is described, permitting a detailed and systematic analysis of the factors governing self-assembly of amphipathic helices, e.g. to a four-helical bundle, a subject of major relevance for tertiary structure formation, protein folding and design. Following the Template Assembled Synthetic Proteins (TASP) approach, helices of different packing potential are competitively assembled in solution with a preformed two-helix TASP molecule, and after equilibration are covalently attached ('template trapping') via chemoselective thioether formation. The quantitative analysis of the individual TASP molecules by high performance liquid chromatography (HPLC) and electrospray mass spectrometry (ES-MS) allows the delineation of the role of complementary packing in helix bundle formation. The procedure established represents a general tool for the experimental verification of modern concepts in molecular recognition.
本文描述了一种实验系统，该系统能够对两亲性螺旋自组装的影响因素进行详细且系统的分析，例如形成四螺旋束，这是与三级结构形成、蛋白质折叠和设计密切相关的主题。按照模板组装合成蛋白（TASP）方法，将具有不同堆积潜力的螺旋与预先形成的双螺旋TASP分子在溶液中竞争性组装，平衡后通过化学选择性硫醚形成进行共价连接（“模板捕获”）。通过高效液相色谱（HPLC）和电喷雾质谱（ES-MS）对各个TASP分子进行定量分析，从而确定互补堆积在螺旋束形成中的作用。所建立的程序是对分子识别现代概念进行实验验证的通用工具。
