Note: Intro -- Glycan-Based Cellular Communication: Techniques for Carbohydrate-Protein Interactions -- ACS Symposium Series1346 -- Glycan-Based Cellular Communication: Techniques for Carbohydrate-Protein Interactions -- Library of Congress Cataloging-in-Publication Data -- Foreword -- Preface -- Recent Advancements in Arrayed Technologies and Emerging Themes in the Identification of Glycan-Protein Interactions -- Insights into Antibody-Carbohydrate Recognition from Neoglycoprotein Microarrays -- Generation of a Heparan Sulfate Mutant Cell Library and Its Application to Determine the Structure-Function Relationship of Heparan Sulfate in Facilitating FGF2-FGFR1 Signaling -- Polyvalent Glycan-Quantum Dots as Multifunctional Structural Probes for Multivalent Lectin-Carbohydrate Interactions -- Automated Identification of Lectin Fine Specificities from Glycan-Array Data -- Label-Free Biosensors for Studying Carbohydrate-Protein Interaction -- Sketching the Glycan Hallmark of Intact Cells Using Lectin Microarray -- Methods to Investigate Innate Immune Receptors and Their Carbohydrate-Based Ligands -- Morphogenesis of the Mammalian Tectorial Membrane: Unveiling the Surface Roles of a Matrix Organizer, Alpha Tectorin -- Mass Spectrometric Analysis of Protein Glycosylation -- Editors' Biographies -- Indexes -- Indexes -- Author Index -- Subject Index -- Preface -- 1 -- Recent Advancements in Arrayed Technologies and Emerging Themes in the Identification of Glycan-Protein Interactions -- Introduction -- Figure 1. Glycoconjugates and glycan-GBP interactions. (A) Glycans can occur as monosaccharides, oligomers, or polymers linked to lipids or proteins, which generate glycolipids or glycoproteins, respectively. (B) Interplay of glycan-GBP interactions in influenza infection. , Figure 2. Facets and challenges of analyzing glycan-GBP interactions. (A) Glycan-GBP interactions are unique due to the ability of glycans to act as a ligand for multiple GBPs, as well as the ability of one GBP to interact with multiple glycans. (B) Glycan-GBP interactions can occur in cis or trans configurations to modulate receptor activity. (C) Different protein glycoforms may interact with different proteins, potentially resulting in varying biological functions. -- Recent Developments and Findings in Glycan and Lectin Microarrays -- Glycan and Lectin Microarray Basics -- Shotgun Glycomics and Beam Search Arrays -- SPR Analyses in Arrays -- Figure 3. Arrayed technologies to survey glycan-GBP interactions and recent improvements. (A) General features of a glycan or lectin array. Shown here are glycans immobilized on a solid surface, and a GBP (yellow circle) tagged with a fluorophore (green star) is incubated with the array. Spot-based fluorescent signals are acquired to then determine glycan binding. (B) Improvements to the classic array using new microfluidic and printing technologies. Reproduced with permission from Reference 32. -- Glycopolymers and Glycodendrimers in Arrays -- DNA Encoding in Glycan Arrays -- Figure 4. DNA-tagged glycan libraries and cell-based arrays. (A) DNA-tagged Next-NGGMs allow the use of next generation sequencing technologies to decode glycans bound to a particular GBP. Reproduced with permission from Reference 51. Copyright 2019 ACS Publications. (B) In an example of a cell-based array, cells were genetically edited by CRISPR/Cas9 techniques to create a library of cells with altered cell surface glycans. The binding of various GBPs, including influenza HA or other microbial -- Cell-Based Arrays -- Non-Arrayed Systems for Analyzing Glycan-GBP Interactions -- Lectin Affinity Chromatography. , Metabolic Labeling and Photo-Crosslinking -- Mass Spectrometry-Based Approaches -- Proteomics -- Figure 5. Non-arrayed systems for probing glycan-GBP interactions. (A) In multi-lectin affinity chromatography, lectins are immobilized onto beads and incubated with mixtures of glycans or glycoproteins. Nonbound components are washed away, and an iterative process yields the determination of the sample's glycoprofile. (B) In the metabolic labeling approach, unnatural monosaccharides (such as those modified with diazirine tags) are incorporated into cellular glycoconjugates. Photoirradiation of -- Glycoproteomics -- Proximity Labeling -- Figure 6. Proximity labeling of Siglec glycoprotein ligands. FLAG-tagged Siglec-Fc fusion proteins are incubated with live cells containing the putative glycan ligand. Then, a HRP-conjugated anti-FLAG antibody is added, to generate Siglec-HRP complexes. Upon the addition of biotin tyramide and hydrogen peroxide, radicalization occurs to label the protein ligands of Siglec. Affinity purification and LC-MS/MS are then carried out to identify the glycoprotein ligands. Reproduced with permission fro -- Outstanding Hurdles, Conclusions, and Outlook for the Future -- References -- 2 -- Insights into Antibody-Carbohydrate Recognition from Neoglycoprotein Microarrays -- Introduction -- Glycan Microarrays -- Figure 1. Profiling carbohydrate-protein interactions with glycan microarrays. -- Neoglycoprotein Microarrays. , Figure 2. Formation of neoglycoproteins. Carbohydrates can be conjugated to proteins using a variety of chemistries. Several examples of conjugation reactions used to form neoglycoproteins are shown. In each case, the sugar is attached to the side chains of lysine residues on the protein. Top: reductive amination of an oligosaccharide lactol to amines on albumin. Middle: EDC activation of a linker with a carboxylic acid. Bottom: Copper-mediated click reaction between a sugar-azide and alkyne-mod -- Figure 3. Representation of a neoglycoprotein. A model of a GD2-BSA neoglycoprotein was generated by attaching 10 GD2 molecules to 10 different lysine residues of bovine serum albumin via a 9-atom linker commonly used by our group. Bovine serum albumin was obtained from the crystal structure 71. All lysine residues are colored blue. No molecular dynamics simulations or minimization has been carried out, This model is for illustrative purposes. -- Analysis of Germline Antibody Binding Properties Using Neoglycoprotein Microarrays -- Figure 4. Glycan structures of GD2 and GQ2. -- Conclusions -- Acknowledgments -- References -- 3 -- Generation of a Heparan Sulfate Mutant Cell Library and Its Application to Determine the Structure-Function Relationship of Heparan Sulfate in Facilitating FGF2-FGFR1 Signaling -- Heparan Sulfate -- HS Structure-Function Study Using HS Mutant Cells. , Figure 1. Generation of an HS mutant MLEC library and coreceptor function of HS in FGF signaling. A. HS biosynthesis and remodeling genes. The genes targeted for the HS mutant cell library are highlighted in red. The pattern of sulfation and epimerization modifications along the chain forms specific binding sites for protein ligands, as highlighted here for FGF, FGFR, and antithrombin. B. Schematic structure of wild-type and mutant HS structures expressed by their corresponding cell lines within -- Application of the HS Mutant Library to Determine the Structural Features Required for HS' Coreceptor Function in FGF2-FGFR1 Signaling -- Acknowledgments -- References -- 4 -- Polyvalent Glycan-Quantum Dots as Multifunctional Structural Probes for Multivalent Lectin-Carbohydrate Interactions -- Introduction -- Lectin Multivalency -- DC-SIGN and DC-SIGNR -- Figure 1. A schematic structure model of DC-SIGN. -- Designing Polyvalent Glycan-Nanoparticle Probes for Lectin-Glycan Interactions -- Figure 2. Schematic depicting the FRET process between an excited glycan-QD and dye-labeled DC-SIGN. -- Glycan-QD Design -- Figure 3. Chemical structure of the DHLA-EGn-glycan multifunctional ligands where n = 3 or 11 and glycan = Man or DiMan (Man-α-1,2-Man). -- Probing Multivalent Lectin-Carbohydrate Interactions Using Glycan-QDs -- Differentiating DC-SIGN/R Binding Modes Using Glycan-QDs -- Figure 4. Background corrected emission spectra obtained for the interaction of (A) DC-SIGN with QD-PEG13-Man -- (B) DC-SIGN with QD-EG3-Man -- (C) DC-SIGNR with QD-PEG13-Man -- (D) DC-SIGNR with QD-EG3-Man. , and (E) DC-SIGN CRD with QD-EG3-Man. The final QD concentration is fixed at 40 nM. (F) Plots of the apparent FRET ratio (I626/I554) versus protein concentration for DC-SIGN/R binding to QD-EG3-Man and QD-PEG13-Man. Reproduced with permission from reference 26. Copyright 2016 Wiley-VCH Verlag GmbH.