Abstract

Pseudomonas aeruginosa is a widespread opportunistic pathogen that is capable of colonizing various human tissues and is resistant to many antibiotics. LecA is a galactose binding tetrameric lectin involved in adhesion, infection and biofilm formation. This study reports on the binding characteristics of mono- and divalent (chelating) ligands to LecA using different techniques. These techniques include affinity capillary electrophoresis, bio-layer interferometry, native mass spectrometry and a thermal shift assay. Aspects of focus include: affinity, selectivity, binding kinetics and residence time. The affinity of a divalent ligand was determined to be in the low-nanomolar range for all of the used techniques and with a ligand residence time of approximately 7 h, while no strong binding was seen to related lectin tetramers. Each of the used techniques provides a unique and complementary insight into the chelation based binding mode of the divalent ligand to the LecA tetramer.

中文翻译：

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使用多种技术评估铜绿假单胞菌凝集素 LecA 与二价半乳糖苷的结合特性

摘要

铜绿假单胞菌是一种广泛存在的机会性病原体，能够定殖各种人体组织并对许多抗生素产生抗药性。LecA 是一种半乳糖结合四聚体凝集素，参与粘附、感染和生物膜形成。本研究报告了使用不同技术对 LecA 的单价和二价（螯合）配体的结合特性。这些技术包括亲和毛细管电泳、生物层干涉法、自然质谱法和热位移测定法。关注的方面包括：亲和力、选择性、结合动力学和停留时间。对于所有使用的技术，二价配体的亲和力被确定为在低纳摩尔范围内，配体停留时间约为 7 小时，而未观察到与相关凝集素四聚体的强结合。