Červený Jakub, Heine Viktoria, Hovorková Michaela, Brož Petr, Filipová Eliška, Kulik Natalia, Hubálek Martin, Cvačka Josef, Petrásková Lucie, Florencio-Zabaleta Mirane, Delgado Sandra, Pelantová Helena, Bosáková Zuzana, Elling Lothar, Jiménez-Barbero Jesus, Ardá Ana, Křen Vladimír, Bojarová Pavla
Institute of Microbiology of the Czech Academy of Sciences, Vídeňská 1083, CZ-142 00 Prague 4, Czech Republic.
Department of Analytical Chemistry, Faculty of Science, Charles University, Hlavova 8, CZ-128 43 Prague 2, Czech Republic.
Biomacromolecules. 2025 Aug 11;26(8):4995-5009. doi: 10.1021/acs.biomac.5c00377. Epub 2025 Jul 7.
Galectins are small human proteins participating in inflammation processes, immune response, and cancerogenesis. Tandem-repeat galectins comprising Gal-4, Gal-8, and Gal-9 are a vital yet less studied part of the galectin fingerprint in cancer-related processes. The present work studies a library of prepared multivalent neo-glycoproteins decorated with poly--acetyllactosamine and human-milk-type oligosaccharides as ligands of this underexplored family of tandem-repeat galectins. A thorough binding evaluation by ELISA and biolayer interferometry was complemented with a detailed epitope mapping both from the galectin and the glycoconjugate viewpoints by nuclear magnetic resonance. The found interactions in the galectin binding site were correlated to data from molecular modeling. The present work reveals pioneer information on the binding of tandem-repeat galectins to multivalent glycoconjugates carrying complex carbohydrate ligands and represents an invaluable starting point for the development of new high-affinity tailored ligands of tandem-repeat galectins, needed both for diagnosis and therapy.
半乳糖凝集素是参与炎症过程、免疫反应和癌症发生的小型人类蛋白质。由Gal-4、Gal-8和Gal-9组成的串联重复半乳糖凝集素是癌症相关过程中半乳糖凝集素指纹图谱的重要组成部分，但研究较少。本研究工作针对一个由多价新糖蛋白组成的文库展开，这些新糖蛋白以聚乙酰乳糖胺和人乳型寡糖作为修饰，作为这个尚未充分研究的串联重复半乳糖凝集素家族的配体。通过酶联免疫吸附测定（ELISA）和生物层干涉术进行了全面的结合评估，并通过核磁共振从半乳糖凝集素和糖缀合物两个角度进行了详细的表位作图。在半乳糖凝集素结合位点发现的相互作用与分子模拟数据相关。本研究工作揭示了关于串联重复半乳糖凝集素与携带复杂碳水化合物配体的多价糖缀合物结合的前沿信息，为开发用于诊断和治疗所需的新型高亲和力定制串联重复半乳糖凝集素配体提供了宝贵的起点。
