Neubacher Saskia, Saya Jordy M, Amore Alessia, Grossmann Tom N
Department of Chemistry & Pharmaceutical Sciences , VU University Amsterdam , De Boelelaan 1083 , 1081 HV Amsterdam , The Netherlands.
J Org Chem. 2020 Feb 7;85(3):1476-1483. doi: 10.1021/acs.joc.9b02490. Epub 2019 Dec 16.
Protein macrocyclization represents a very efficient strategy to increase the stability of protein tertiary structures. Here, we describe a panel of novel C3-symmetric tris-electrophilic agents and their use for the cyclization of proteins. These electrophiles are reacted with a protein domain harboring three solvent-exposed cysteine residues, resulting in the in situ cyclization of the protein (INCYPRO). We observe a clear dependency of cross-linking rates on the electrophilicity. All nine obtained cross-linked protein versions show considerably increased thermal stability (up to 29 °C increased melting temperature) when compared to that of the linear precursor. Most interestingly, the degree of stabilization correlates with the hydrophilicity of the cross-link. These results will support the development of novel cross-linked proteins and enable a more rational design process.
蛋白质大环化是提高蛋白质三级结构稳定性的一种非常有效的策略。在此，我们描述了一组新型的C3对称三亲电试剂及其在蛋白质环化中的应用。这些亲电试剂与含有三个溶剂暴露半胱氨酸残基的蛋白质结构域反应，导致蛋白质原位环化（INCYPRO）。我们观察到交联速率明显依赖于亲电性。与线性前体相比，所有九个获得的交联蛋白变体均表现出显著提高的热稳定性（熔解温度提高高达29°C）。最有趣的是，稳定程度与交联的亲水性相关。这些结果将支持新型交联蛋白的开发，并实现更合理的设计过程。
