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Design of Thermoresponsive Elastin-Like Glycopolypeptides for Selective Lectin Binding and Sorting.
Biomacromolecules ( IF 5.5 ) Pub Date : 2020-05-07 , DOI: 10.1021/acs.biomac.0c00374 Lourdes Mónica Bravo Anaya 1 , Rosine Petitdemange 1, 2 , Marie Rosselin 1 , Emmanuel Ibarboure 1 , Bertrand Garbay 1 , Elisabeth Garanger 1 , Timothy J Deming 2 , Sebastien Lecommandoux 1
Biomacromolecules ( IF 5.5 ) Pub Date : 2020-05-07 , DOI: 10.1021/acs.biomac.0c00374 Lourdes Mónica Bravo Anaya 1 , Rosine Petitdemange 1, 2 , Marie Rosselin 1 , Emmanuel Ibarboure 1 , Bertrand Garbay 1 , Elisabeth Garanger 1 , Timothy J Deming 2 , Sebastien Lecommandoux 1
Affiliation
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Selective lectin binding and sorting was achieved using thermosensitive glycoconjugates derived from recombinant elastin-like polypeptides (ELPs) in simple centrifugation-precipitation assays. A recombinant ELP, (VPGXG)40, containing periodically spaced methionine residues was used to enable chemoselective postsynthetic modification via thioether alkylation using alkyne functional epoxide derivatives. The resulting sulfonium groups were selectively demethylated to give alkyne functionalized homocysteine residues, which were then reacted with azido-functionalized monosaccharides to obtain ELP glycoconjugates with periodic saccharide functionality. These modifications were also found to allow modulation of ELP temperature dependent water solubility. The multivalent ELP glycoconjugates were evaluated for specific recognition, binding and separation of the lectin Ricinus communis agglutinin (RCA120) from a complex protein mixture. RCA120 and ELP glycoconjugate interactions were evaluated using laser scanning confocal microscopy and dynamic light scattering. Due to the thermoresponsive nature of the ELP glycoconjugates, it was found that heating a mixture of galactose-functionalized ELP and RCA120 in complex media selectively yielded a phase separated pellet of ELP-RCA120 complexes. Based on these results, ELP glycoconjugates show promise as designer biopolymers for selective protein binding and sorting.
中文翻译：
用于选择性凝集素结合和分选的热反应性类似弹性蛋白的糖多肽的设计。
在简单的离心沉淀试验中，使用衍生自重组弹性蛋白样多肽（ELP）的热敏糖缀合物实现了选择性凝集素的结合和分选。重组ELP（VPGXG）40含有周期性间隔的甲硫氨酸残基的H2O3，被用于通过炔烃功能性环氧化物衍生物通过硫醚烷基化进行化学选择性的合成后修饰。将产生的de基团选择性地去甲基化，得到炔基官能化的高半胱氨酸残基，然后使其与叠氮基官能化的单糖反应以获得具有周期性糖官能度的ELP糖缀合物。还发现这些修饰可以调节依赖于ELP温度的水溶性。评价了多价ELP糖缀合物从复杂蛋白混合物中对凝集素Ricinus communis凝集素（RCA 120）的特异性识别，结合和分离。RCA 120使用激光扫描共聚焦显微镜和动态光散射评估了ELP和ELP糖缀合物之间的相互作用。由于ELP糖缀合物的热响应性，发现在复合介质中加热半乳糖官能化的ELP和RCA 120的混合物选择性地产生了ELP-RCA 120复合物的相分离的沉淀。基于这些结果，ELP糖缀合物显示出作为设计者生物聚合物用于选择性蛋白质结合和分选的前景。
更新日期：2020-05-07
中文翻译：
用于选择性凝集素结合和分选的热反应性类似弹性蛋白的糖多肽的设计。
在简单的离心沉淀试验中，使用衍生自重组弹性蛋白样多肽（ELP）的热敏糖缀合物实现了选择性凝集素的结合和分选。重组ELP（VPGXG）40含有周期性间隔的甲硫氨酸残基的H2O3，被用于通过炔烃功能性环氧化物衍生物通过硫醚烷基化进行化学选择性的合成后修饰。将产生的de基团选择性地去甲基化，得到炔基官能化的高半胱氨酸残基，然后使其与叠氮基官能化的单糖反应以获得具有周期性糖官能度的ELP糖缀合物。还发现这些修饰可以调节依赖于ELP温度的水溶性。评价了多价ELP糖缀合物从复杂蛋白混合物中对凝集素Ricinus communis凝集素（RCA 120）的特异性识别，结合和分离。RCA 120使用激光扫描共聚焦显微镜和动态光散射评估了ELP和ELP糖缀合物之间的相互作用。由于ELP糖缀合物的热响应性，发现在复合介质中加热半乳糖官能化的ELP和RCA 120的混合物选择性地产生了ELP-RCA 120复合物的相分离的沉淀。基于这些结果，ELP糖缀合物显示出作为设计者生物聚合物用于选择性蛋白质结合和分选的前景。
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