Kurhade Suresh E, Weiner Jack D, Gao Fei Philip, Farrell Mark P
Department of Medicinal Chemistry, The University of Kansas, 2034 Becker Drive, Lawrence, KS, 66047, USA.
Protein Production Group, The University of Kansas, 2034 Becker Drive, Lawrence, KS, 66047, USA.
Angew Chem Int Ed Engl. 2021 May 25;60(22):12313-12318. doi: 10.1002/anie.202101249. Epub 2021 Apr 26.
An engineered cyanovirin-N homologue that exhibits specificity for high mannose N-glycans has been constructed to aid type I α 1,2-mannosidase inhibitor discovery and development. Engineering the lectins C-terminus permitted facile functionalization with fluorophores via a sortase and click strategy. The resulting lectin constructs exhibit specificity for cells presenting high mannose N-glycans. Importantly, these lectin constructs can also be applied to specifically assess changes in cell surface glycosylation induced by type I mannosidase inhibitors. Testing the utility of these lectin constructs led to the discovery of type I mannosidase inhibitors with nanomolar potency. Cumulatively, these findings reveal the specificity and utility of the functionalized cyanovirin-N homologue constructs, and highlight their potential in analytical contexts that require high mannose-specific lectins.
为了辅助I型α 1,2-甘露糖苷酶抑制剂的发现与开发，构建了一种对高甘露糖型N-聚糖具有特异性的工程化氰病毒素-N同源物。对凝集素的C端进行工程改造，使其能够通过分选酶和点击策略方便地与荧光团进行功能化修饰。所得的凝集素构建体对呈现高甘露糖型N-聚糖的细胞具有特异性。重要的是，这些凝集素构建体还可用于特异性评估I型甘露糖苷酶抑制剂诱导的细胞表面糖基化变化。对这些凝集素构建体的实用性进行测试，发现了具有纳摩尔效力的I型甘露糖苷酶抑制剂。总体而言，这些发现揭示了功能化氰病毒素-N同源物构建体的特异性和实用性，并突出了它们在需要高甘露糖特异性凝集素的分析环境中的潜力。
