Automated Glycan Assembly of ¹⁹F‐labeled Glycan Probes Enables High‐Throughput NMR Studies of Protein–Glycan Interactions

Abstract

Protein-glycan interactions mediate important biological processes,i ncluding pathogen host invasion and cellular communication. Herein, we showcase an expedite approach that integrates automated glycan assembly (AGA) of 19 F-labeled probes and high-throughput NMR methods,e nabling the study of protein-glycan interactions.Synthetic Lewis type 2a ntigens were screened against seven glycan binding proteins (GBPs), including DC-SIGN and BambL, respectively involved in HIV-1 and lung infections in immunocompromised patients,c onfirming the preference for fucosylated glycans (Le x ,H type 2, Le y ). Previously unknown glycanlectin weak interactions were detected, and thermodynamic data were obtained. Enzymatic reactions were monitored in real-time,d elivering kinetic parameters.T hese results demonstrate the utility of AGAc ombined with 19 FNMR for the discovery and characterization of glycan-protein interactions, opening up new perspectives for 19 F-labeled complex glycans.

The Flexibility of Oligosaccharides Unveiled Through Residual Dipolar Coupling Analysis

“…NMR has been extensively employed to assess the conformational, dynamic and recognition features of these flexible molecules. Recent developments using paramagnetic NMR approaches ( Suzuki et al, 2017 ; Fernández de Toro et al, 2018 ) or NMR-active nuclei ( 13 C, 19 F) as labels permitted to circumvent the tremendous overlapping problem inherent to glycans ( Fittolani et al, 2021 ; Moure et al, 2021 ), especially in the case of homo-oligosaccharides.…”

Riclinoctaose Attenuates Renal Ischemia-Reperfusion Injury by the Regulation of Macrophage Polarization

“…The interfering of riclinoctaose/ mDC-SIGN blocked the function of riclinoctaose on M2 polarization, consequently impairing the renoprotective effect of riclinoctaose. hDC-SIGN was reported to be highly expressed on the population of myeloid cells (Geijtenbeek al., 2000), and it has turned out to be a target for developing antiviral drugs since it can interact with viral glycans to facilitate virus spreading and exacerbates inflammatory reactions (Jarvis et al, 2019;Fittolani et al, 2021;Ramos-Soriano and Rojo, 2021). hDC-SIGN has been found to be involved in pattern recognition of a broad range of pathogen-derived ligands, meditation of intercellular adhesion, and self-glycoproteins receptor (van Kooyk and Geijtenbeek, 2003;Garcia-Vallejo and van Kooyk, 2013;Jarvis et al, 2019).…”

“…hDC-SIGN has been found to be involved in pattern recognition of a broad range of pathogen-derived ligands, meditation of intercellular adhesion, and self-glycoproteins receptor (van Kooyk and Geijtenbeek, 2003;Garcia-Vallejo and van Kooyk, 2013;Jarvis et al, 2019). Recently, hDC-SIGN has gained considerable attention for its participation in the infection process of several pathogens (Fittolani et al, 2021;Ramos-Soriano and Rojo, 2021). Mouse DC-SIGN is one of a family of C-type lectin genes syntenic and homologous to human DC-SIGN.…”

Allosterische, Wirkstoff‐zugängliche Bindestellen in β‐Propeller‐Lektinen

“…Im Vergleich zu BambL‐WT behielt T18S seine Affinität für 2FF bei (Abbildung 4 e, WT: K d =7,9±0,2 μM, T18S: K d =8,2±0,2 μM). BambL‐T18S zeigte jedoch eine fast zweifach geringere Affinität für F‐H Typ 2 (Abbildung S17 b, K d =16,7±2,5 μM) als BambL‐WT ( K d =9±2 μM [28] ). Um die Bindung von F‐H Typ 2 an 15 N‐BambL‐T18S zu überprüfen, verwendeten wir TROSY‐NMR und verglichen die CSPs mit dem WT‐Protein (Abbildung S17 c).…”