Cristina Di Carluccio, Linda Cerofolini, Miguel Moreira, Frédéric Rosu, Luis Padilla-Cortés, Giulia Roxana Gheorghita, Zhuojia Xu, Abhishek Santra, Hai Yu, Shinji Yokoyama, Taylor E. Gray, Chris D. St. Laurent, Yoshiyuki Manabe, Xi Chen, Koichi Fukase, Matthew S. Macauley, Antonio Molinaro, Tiehai Li, Barbara A. Bensing, Roberta Marchetti*, Valérie Gabelica, Marco Fragai and Alba Silipo*,
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Despite a conserved overall domain organization of SRR adhesins, the Siglec-like binding regions (SLBRs) are highly variable, affecting the recognition of a wide range of sialoglycans. SLBR-N from the SRR glycoprotein of <i>S. gordonii</i> UB10712 possesses the remarkable ability to recognize complex core 2 <i>O</i>-glycans. We here employed a multidisciplinary approach, including flow cytometry, native mass spectrometry, isothermal titration calorimetry, NMR spectroscopy from both protein and ligand perspectives, and computational methods, to investigate the ligand specificity and binding preferences of SLBR-N when interacting with mono- and disialylated core 2 <i>O</i>-glycans. We determined the means by which SLBR-N preferentially binds branched α2,3-disialylated core 2 <i>O</i>-glycans: a selected conformation of the 3′SLn branch is accommodated into the main binding site, driving the sTa branch to further interact with the protein. 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引用次数: 0
摘要
戈登链球菌(Streptococcus gordonii)是一种革兰氏阳性细菌,通常在人的口腔中定植,但也可引起局部或全身性疾病。暴露在戈登链球菌表面的富丝氨酸重复(SRR)糖蛋白与人类唾液、血浆和血小板糖蛋白上的糖基化聚糖结合,这可能会导致口腔定植和心内膜感染。尽管 SRR 粘附蛋白的整体结构域是保守的,但 Siglec 样结合区(SLBR)的变化很大,影响了对多种聚糖的识别。来自 S. gordonii UB10712 的 SRR 糖蛋白的 SLBR-N 具有识别复杂的核心 2 O-聚糖的非凡能力。在此,我们采用了一种多学科方法,包括流式细胞术、原生质谱法、等温滴定量热法、从蛋白质和配体角度进行的核磁共振波谱分析以及计算方法,来研究 SLBR-N 与单酰化和二酰化核心 2 O-聚糖相互作用时的配体特异性和结合偏好。我们确定了 SLBR-N 优先结合支化的 α2,3-二氨酰化核心 2 O-聚糖的方式:3′SLn 支的选定构象被容纳到主结合位点,驱动 sTa 支进一步与蛋白质相互作用。与此同时,SLBR-N 在聚糖结合袋的 CD 环上呈现开放构象,从而可以容纳整个复合核心 2 O-聚糖。这些发现为开发新型工具检测特定的复合 O 型糖结构奠定了基础,并为设计和开发潜在的链球菌感染治疗药物铺平了道路。
Molecular Insights into O-Linked Sialoglycans Recognition by the Siglec-Like SLBR-N (SLBRUB10712) of Streptococcus gordonii
Streptococcus gordonii is a Gram-positive bacterial species that typically colonizes the human oral cavity, but can also cause local or systemic diseases. Serine-rich repeat (SRR) glycoproteins exposed on the S. gordonii bacterial surface bind to sialylated glycans on human salivary, plasma, and platelet glycoproteins, which may contribute to oral colonization as well as endocardial infections. Despite a conserved overall domain organization of SRR adhesins, the Siglec-like binding regions (SLBRs) are highly variable, affecting the recognition of a wide range of sialoglycans. SLBR-N from the SRR glycoprotein of S. gordonii UB10712 possesses the remarkable ability to recognize complex core 2 O-glycans. We here employed a multidisciplinary approach, including flow cytometry, native mass spectrometry, isothermal titration calorimetry, NMR spectroscopy from both protein and ligand perspectives, and computational methods, to investigate the ligand specificity and binding preferences of SLBR-N when interacting with mono- and disialylated core 2 O-glycans. We determined the means by which SLBR-N preferentially binds branched α2,3-disialylated core 2 O-glycans: a selected conformation of the 3′SLn branch is accommodated into the main binding site, driving the sTa branch to further interact with the protein. At the same time, SLBR-N assumes an open conformation of the CD loop of the glycan-binding pocket, allowing one to accommodate the entire complex core 2 O-glycan. These findings establish the basis for the generation of novel tools for the detection of specific complex O-glycan structures and pave the way for the design and development of potential therapeutics against streptococcal infections.
A multidisciplinary approach combining NMR spectroscopy, native mass spectrometry, flow cytometry, ITC and MD simulation unveiled the binding details of core 2 O-glycans to the Siglec-like adhesin SLBR-N of S. gordonii.
期刊介绍:
ACS Central Science publishes significant primary reports on research in chemistry and allied fields where chemical approaches are pivotal. As the first fully open-access journal by the American Chemical Society, it covers compelling and important contributions to the broad chemistry and scientific community. "Central science," a term popularized nearly 40 years ago, emphasizes chemistry's central role in connecting physical and life sciences, and fundamental sciences with applied disciplines like medicine and engineering. The journal focuses on exceptional quality articles, addressing advances in fundamental chemistry and interdisciplinary research.