Perfluorophenyl Azide Immobilization Chemistry for Single Molecule Force Spectroscopy of the Concanavalin A/Mannose Interaction

Madwar, Carolin; Kwan, William Chu; Deng, Lih‐Wen; Ramström, Olof; Schmidt, Rolf; Zou, Shan; Cuccia, Louis A.

doi:10.1021/la1036579

Cited by 9 publications

(8 citation statements)

References 32 publications

(66 reference statements)

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“…Frequencies of adhesive events at tip–surface separation larger than 50 nm for the different glycomacromolecules are shown as histograms in Figure 4 . We find that dissociation forces occur most frequently in the range of 50–90 pN, which is similar to earlier studies on the interaction between mono‐ and multivalent Man ligands with ConA . For oligomeric ligands, the dissociation force increases with increasing Man valency, where the most significant increase occurs between Man‐2 (57 ± 19 pN) and Man‐3 (72 ± 20 pN).…”

Section: Resultssupporting

confidence: 88%

Multivalent Interactions of Polyamide Based Sequence‐Controlled Glycomacromolecules with Concanavalin A

Calle

Gerke

Chang

et al. 2019

Macromolecular Bioscience

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Binding of mannose presenting macromolecules to the protein receptor concanavalin A (ConA) is investigated by means of single‐molecule atomic force spectroscopy (SMFS) in combination with dynamic light scattering and molecular modeling. Oligomeric (Mw ≈ 1.5–2.5 kDa) and polymeric (Mw ≈ 22–30 kDa) glycomacromolecules with controlled number and positioning of mannose units along the scaffolds accessible by combining solid phase synthesis and thiol–ene coupling are used as model systems to assess the molecular mechanisms that contribute to multivalent ConA–mannose complexes. SMFS measurements show increasing dissociation force from monovalent (≈57 pN) to pentavalent oligomers (≈75 pN) suggesting subsite binding to ConA. Polymeric glycomacromolecules with larger hydrodynamic diameters compared to the binding site spacing of ConA exhibit larger dissociation forces (≈80 pN), indicating simultaneous dissociation from multiple ConA binding sites. Nevertheless, although simultaneous dissociation of multiple ligands could be expected for such multivalent systems, predominantly single dissociation events are observed. This is rationalized by strong coiling of the macromolecules' polyamide backbone due to intramolecular hydrogen bonding hindering unfolding of the coil. Therefore, this study shows that the design of glycopolymers for multivalent receptor binding and clustering must consider 3D structure and intramolecular interactions of the scaffold.

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Section: Resultssupporting

confidence: 88%

Multivalent Interactions of Polyamide Based Sequence‐Controlled Glycomacromolecules with Concanavalin A

Calle

Gerke

Chang

et al. 2019

Macromolecular Bioscience

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“…Several methods to study carbohydrate–protein interactions can be employed to address this difference, e.g., isothermal titration calorimetry (ITC) [44], hemagglutination inhibition [40], precipitation inhibition [45], equilibrium dialysis [46], spectrophotometry [47], enzyme-linked immunosorbent assay (ELISA) [48], fluorescence assay [49,50], nuclear magnetic resonance (NMR) [51], surface plasmon resonance (SPR) [41,52], quartz-crystal microbalance (QCM) [53,54], carbohydrate microarrays [55], and scanning probe microscopy (SPM) [56]. Of these, two major methods are especially advantageous: ITC and solid-phase binding techniques, the combination of which results in elucidation of overall binding performances.…”

Section: Introductionmentioning

confidence: 99%

“…Solid-supported techniques are thus playing crucial roles for studies of various carbohydrate–protein interactions, and as a result, methods for immobilization of carbohydrates on surfaces and resins have become an important research topic. In this context, the well-established perfluoro-phenylazide (PFPA) chemistry enables one-step, facile, and robust attachment of organic molecules on solid surfaces, by either thermal- or photo-activation, resulting in versatile surface modification for a wide range of applications [41,49,52–56,61–64]. …”

Section: Introductionmentioning

confidence: 99%

Stereocontrolled 1-S-glycosylation and comparative binding studies of photoprobe-thiosaccharide conjugates with their O-linked analogs

Deng

Wang

Uppalapati

et al. 2013

Pure and Applied Chemistry

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The use of thioglycosides and other glycan derivatives with anomeric sulfur linkages is gaining increasing interest, both in synthesis and in various biological contexts. Herein, we demonstrate the occurrence and circumvention of anomerization during 1-S-glycosylation reactions, and present highly efficient and stereocontrolled syntheses of a series of photoprobe-thiosaccharide conjugates. Mutarotation of glycosyl thiols proved to be the origin of the anomeric mixtures formed, and kinetic effects could be used to circumvent anomerization. The synthesized carbohydrate conjugates were then evaluated by both solution- and solid-phase-based techniques. Both binding results showed that the S-linked glyco-sides interact with their cognate lectins comparably to the corresponding O-analogs in the present cases, thus demonstrating the reliability of the solid-support platform built upon our photo-initiated carbohydrate immobilization method for probing protein bindings, and showing the potential of combining these two means for studying carbohydrate–protein interactions.

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“…The mannobiose was anchored to gold surfaces via a linker molecule (ATFMB or 4-azido-2,3,5,6-tetrafluoro- N -(2-mercaptoethyl)benzamide) that has a thiol functional group at one end and an aryl azide on the other. The linkers self-assemble on gold surfaces via the thiol end, and the sugar residues were photochemically linked to the aryl azide end. , …”

Section: Resultsmentioning

confidence: 99%

Mannose Surfaces Exhibit Self-Latching, Water Structuring, and Resilience to Chaotropes: Implications for Pathogen Virulence

Abeyratne-Perera

Chandran

2017

Langmuir

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Several viral and fungal pathogens, including HIV, SARS, Dengue, Ebola, and Cryptococcus neoformans, display a preponderance of mannose residues on their surface, particularly during the infection cycle or in harsh environments. The innate immune system, on the other hand, abounds in mannose receptors which recognize mannose residues on pathogens and trigger their phagocytosis. We pose the question if there is an advantage for pathogens to display mannose on their surface, despite these residues being recognized by the immune system. The surface properties and interactions of opposing monolayers of mannobiose (disaccharide of mannose) were probed using atomic force spectroscopy. Unlike its diastereoisomer lactose, mannobiose molecules exhibited lateral packing interactions that manifest on the surface scale as a self-recognizing latch. A break-in force is required for opposing surfaces to penetrate and a breakout (or self-adhesion force) of similar magnitude is required for penetrated surfaces to separate. A hierarchy of self-adhesion forces was distinguished as occurring at the single residue (∼25 pN), cluster (∼250 pN), monolayer (∼1.1 nN), and supramonolayer level. The break-in force and break-out force appear resilient to the presence of simple chaotropes that attenuate a layer of structured water around the mannose surface. The layer of structured water otherwise extends to distances several times longer than a mannobiose residue, indicating a long-range propagation of the hydrogen bonding imposed by the residues. The span of the structured water increases with the velocity of an approaching surface, similar to shear thickening, but fissures at higher approach velocities. Our studies suggest that mannose residues could guide interpathogen interactions, such as in biofilms, and serve as a moated fortress for pathogens to hide behind to resist detection and harsh environments.

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Perfluorophenyl Azide Immobilization Chemistry for Single Molecule Force Spectroscopy of the Concanavalin A/Mannose Interaction

Cited by 9 publications

References 32 publications

Multivalent Interactions of Polyamide Based Sequence‐Controlled Glycomacromolecules with Concanavalin A

Multivalent Interactions of Polyamide Based Sequence‐Controlled Glycomacromolecules with Concanavalin A

Stereocontrolled 1-S-glycosylation and comparative binding studies of photoprobe-thiosaccharide conjugates with their O-linked analogs

Mannose Surfaces Exhibit Self-Latching, Water Structuring, and Resilience to Chaotropes: Implications for Pathogen Virulence

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