Department of Chemistry, University of British Columbia, Vancouver, British Columbia V6T 1Z1, Canada.
Langmuir. 2011 May 17;27(10):5713-8. doi: 10.1021/la200915d. Epub 2011 Apr 12.
Constructing polyproteins consisting of identical tandem repeats of proteins provides an unambiguous method of investigating the mechanical properties of proteins at the single-molecule level using force spectroscopy techniques. Here we report a maleimide-thiol coupling-based facile method of constructing polyproteins for single-molecule force spectroscopy studies on the mechanical properties of proteins. This method allows for the construction of polyproteins in an efficient fashion under room temperature. The resultant thioether bonds are resistant to reduction and make it possible to carry out single-molecule force spectroscopy studies under various redox conditions. This novel method complements existing polyprotein engineering methods and can be easily applied to a wide variety of proteins.
构建由蛋白质的相同串联重复组成的多聚蛋白，为使用力谱技术在单分子水平上研究蛋白质的机械性质提供了一种明确的方法。在这里，我们报告了一种基于马来酰亚胺-巯基偶联的简便方法，用于构建多聚蛋白，以便在机械性质的单分子力谱研究中使用。该方法允许在室温下以有效的方式构建多聚蛋白。所得的硫醚键对还原具有抗性，使得可以在各种氧化还原条件下进行单分子力谱研究。这种新方法补充了现有的多聚蛋白工程方法，并且可以很容易地应用于各种蛋白质。
