Macromolecular assemblies of complex polysaccharides with galectin-3 and their synergistic effects on function

2017 ◽  
Vol 474 (22) ◽  
pp. 3849-3868 ◽  
Author(s):  
Tao Zhang ◽  
Michelle C. Miller ◽  
Yi Zheng ◽  
Zhongyu Zhang ◽  
Huiting Xue ◽  
...  
Keyword(s):  
Quantum Coherence ◽  
Synergistic Effects ◽  
Lectin Binding ◽  
Model Systems ◽  
Nmr Studies ◽  
Macromolecular Assemblies ◽  
T Cell Apoptosis ◽  
Large Size ◽  
Kd Values ◽  
Galectin 3

Although pectin-derived polysaccharides can antagonize galectin function in various pathological disorders, the nature of their binding interactions needs to be better defined for developing them as drugs. Moreover, given their relatively large size and complexity, pectin-derived polysaccharides are also useful as model systems to assess inter-polysaccharide and protein–polysaccharide interactions. Here, we investigated interactions between galectin-3 (Gal-3) and pectin-derived polysaccharides: a rhamnogalacturonan (RG) and two homogalacturonans (HGs). BioLayer Interferometry and fluorescence-linked immunosorbent assays indicate that these polysaccharides bind Gal-3 with macroscopic or apparent KD values of 49 nM, 46 µM, and 138 µM, respectively. 15N-1H heteronuclear single quantum coherence (HSQC) NMR studies reveal that these polysaccharides interact primarily with the F-face of the Gal-3 carbohydrate recognition domain. Even though their binding to Gal-3 does not inhibit Gal-3-mediated T-cell apoptosis and only weakly attenuates hemagglutination, their combination in specific proportions increases activity synergistically along with avidity for Gal-3. This suggests that RG and HG polysaccharides act in concert, a proposal supported by polysaccharide particle size measurements and 13C-1H HSQC data. Our model has HG interacting with RG to promote increased avidity of RG for Gal-3, likely by exposing additional lectin-binding sites on the RG. Overall, the present study contributes to our understanding of how complex HG and RG polysaccharides interact with Gal-3.

scholarly journals Cross-Linking Effects Dictate the Preference of Galectins to Bind LacNAc-Decorated HPMA Copolymers

2021 ◽  
Vol 22 (11) ◽  
pp. 6000
Author(s):  
Sara Bertuzzi ◽  
Ana Gimeno ◽  
Ane Martinez-Castillo ◽  
Marta G. Lete ◽  
Sandra Delgado ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Lectin Binding ◽  
Building Blocks ◽  
Cryo Electron Microscopy ◽  
Hpma Copolymers ◽  
Hydroxypropyl Methacrylamide ◽  
Galectin 3 ◽  
Nmr Methods ◽  
The Individual ◽  
Statistical Effects

The interaction of multi-LacNAc (Galβ1-4GlcNAc)-containing N-(2-hydroxypropyl) methacrylamide (HPMA) copolymers with human galectin-1 (Gal-1) and the carbohydrate recognition domain (CRD) of human galectin-3 (Gal-3) was analyzed using NMR methods in addition to cryo-electron-microscopy and dynamic light scattering (DLS) experiments. The interaction with individual LacNAc-containing components of the polymer was studied for comparison purposes. For Gal-3 CRD, the NMR data suggest a canonical interaction of the individual small-molecule bi- and trivalent ligands with the lectin binding site and better affinity for the trivalent arrangement due to statistical effects. For the glycopolymers, the interaction was stronger, although no evidence for forming a large supramolecule was obtained. In contrast, for Gal-1, the results indicate the formation of large cross-linked supramolecules in the presence of multivalent LacNAc entities for both the individual building blocks and the polymers. Interestingly, the bivalent and trivalent presentation of LacNAc in the polymer did not produce such an increase, indicating that the multivalency provided by the polymer is sufficient for triggering an efficient binding between the glycopolymer and Gal-1. This hypothesis was further demonstrated by electron microscopy and DLS methods.

NMR studies on Fraser fir Abies fraseri (Pursh) Poir. lignins

Holzforschung ◽  
2005 ◽  
Vol 59 (5) ◽  
pp. 488-496 ◽  
Author(s):  
Mikhail Yu. Balakshin ◽  
Ewellyn A. Capanema ◽  
Barry Goldfarb ◽  
John Frampton ◽  
John F. Kadla
Keyword(s):  
Quantum Coherence ◽  
Lignin Content ◽  
Lower Amount ◽  
Multiple Quantum ◽  
Normal Wood ◽  
Nmr Studies ◽  
Oh Groups ◽  
Fraser Fir ◽  
Heteronuclear Multiple Quantum Coherence ◽  
Balsam Woolly Adelgid

Abstract The composition of mature, juvenile uninfested and juvenile infested Fraser fir wood (Rotholz) and the structures of lignins isolated from these woods were elucidated to verify differences between juvenile and mature wood and the effect of balsam woolly adelgid (BWA) infestation. Milled wood lignin (MWL) isolated from mature, juvenile and Rotholz wood were comprehensively characterized using heteronuclear multiple quantum coherence (HMQC) and quantitative 13C NMR techniques. The Rotholz wood was found to have ∼13% higher lignin content and more than five-fold the amount of galactans than that of the uninfested wood. Rotholz lignin possesses higher amounts of p-hydroxyphenyl units and aliphatic OH groups and a lower amount of alkyl-O-alkyl linkages and dibenzodioxocin moieties. The degree of condensation of the Rotholz lignin was rather similar to that of normal wood. Only small differences in the structure of mature and juvenile wood components were found.

scholarly journals Clicked and long spaced galactosyl- and lactosylcalix[4]arenes: new multivalent galectin-3 ligands

2014 ◽  
Vol 10 ◽  
pp. 1672-1680 ◽  
Author(s):  
Silvia Bernardi ◽  
Paola Fezzardi ◽  
Gabriele Rispoli ◽  
Stefania E Sestito ◽  
Francesco Peri ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Ethylene Glycol ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Lectin Binding ◽  
The Other ◽  
Isomeric Form ◽  
Synthetic Pathway ◽  
Cuaac Reaction ◽  
Galectin 3

Four novel calix[4]arene-based glycoclusters were synthesized by conjugating the saccharide units to the macrocyclic scaffold using the CuAAC reaction and using long and hydrophilic ethylene glycol spacers. Initially, two galactosylcalix[4]arenes were prepared starting from saccharide units and calixarene cores which differ in the relative dispositions of the alkyne and azido groups. Once the most convenient synthetic pathway was selected, two further lactosylcalix[4]arenes were obtained, one in the cone, the other one in the 1,3-alternate structure. Preliminary studies of the interactions of these novel glycocalixarenes with galectin-3 were carried out by using a lectin-functionalized chip and surface plasmon resonance. These studies indicate a higher affinity of lactosyl- over galactosylcalixarenes. Furthermore, we confirmed that in case of this specific lectin binding the presentation of lactose units on a cone calixarene is highly preferred with respect to its isomeric form in the 1,3-alternate structure.

Two-dimensional NMR studies of acrylate copolymers

2009 ◽  
Vol 81 (3) ◽  
pp. 389-415 ◽  
Author(s):  
A. S. Brar ◽  
Ashok Kumar Goyal ◽  
Sunita Hooda
Keyword(s):  
Methyl Methacrylate ◽  
Methyl Acrylate ◽  
Quantum Coherence ◽  
Multiple Bond ◽  
Microstructural Analysis ◽  
2D Nmr ◽  
Correlation Spectroscopy ◽  
Nmr Studies ◽  
Poly Methyl Methacrylate ◽  
Nmr Study

High-resolution NMR spectroscopy is the most versatile, reliable, and generally acceptable technique for the determination of the microstructure of polymers. 2D NMR techniques, along with 1D NMR, have more potential to study absolute configurational assignments and sequence distribution of copolymers. Physical and chemical properties of polymers are influenced fundamentally by their microstructure. We discuss the detailed microstructure analysis of a large number of homopolymers, copolymers, and terpolymers. 2D NMR study of poly(methyl methacrylate) (PMMA), poly(methyl acrylate) (PMA), and poly(methacrylonitrile) (PMAN) is discussed in this article. In addition to homopolymers, 2D heteronuclear single-quantum coherence (HSQC), total correlation spectroscopy (TOCSY), and heteronuclear multiple-bond correlation (HMBC) study of different copolymers such as poly(methyl methacrylate-co-methyl acrylate), poly(styrene-co-methyl methacrylate), and poly(methyl methacrylate-co-methacrylonitrile) have also been reported here. This in turn helps in microstructural analysis of terpolymers such as poly(methacrylonitrile-co-styrene-co-methyl methacrylate), poly(acrylonitrile-co-methyl methacrylate-co-methyl acrylate), and poly(ethylene-co-vinyl acetate-co-carbon monoxide).

scholarly journals Dissecting the Fine Details of Assembly of aT = 3 Phage Capsid

2005 ◽  
Vol 6 (2) ◽  
pp. 119-125 ◽  
Author(s):  
P. G. Stockley ◽  
A. E. Ashcroft ◽  
S. Francese ◽  
G. S. Thompson ◽  
N. A. Ranson ◽  
...  
Keyword(s):  
Rna Binding ◽  
Protein Complexes ◽  
Model Systems ◽  
Replicase Gene ◽  
Nmr Studies ◽  
Partial Explanation ◽  
Stem Loop ◽  
Assembly Pathway

The RNA bacteriophages represent ideal model systems in which to probe the detailed assembly pathway for the formation of aT = 3 quasi-equivalent capsid. For MS2, the assembly reaction can be probedin vitrousing acid disassembled coat protein subunits and a short (19 nt) RNA stem-loop that acts as the translational operator of the replicase gene and leads to sequence-specific sequestration and packaging of the cognate phage RNAin vivo. Reassembly reactions can be initiated by mixing these components at neutral pH. The molecular basis of the sequence-specific RNA–protein interaction is now well understood. Recent NMR studies on the protein demonstrate extensive mobility in the loops of the polypeptide that alter their conformations to form the quasi-equivalent conformers of the final capsid. It seems reasonable to assume that RNA binding results in reduction of this flexibility. However, mass spectrometry suggests that these RNA–protein complexes may only provide one type of quasi-equivalent capsid building block competent to form five-fold axes but not the full shell. Work with longer RNAs suggests that the RNA may actively template the assembly pathway providing a partial explanation of how conformers are selected in the growing shell.

NMR Studies of Coupled Low- and High-Barrier Hydrogen Bonds in Pyridoxal-5‘-phosphate Model Systems in Polar Solution

2007 ◽  
Vol 129 (19) ◽  
pp. 6313-6327 ◽  
Author(s):  
Shasad Sharif ◽  
Gleb S. Denisov ◽  
Michael D. Toney ◽  
Hans-Heinrich Limbach
Keyword(s):  
Hydrogen Bonds ◽  
Model Systems ◽  
Nmr Studies ◽  
Polar Solution

scholarly journals Spatially Selective Heteronuclear Multiple-Quantum Coherence Spectroscopy for Biomolecular NMR Studies

ChemPhysChem ◽  
2014 ◽  
Vol 15 (9) ◽  
pp. 1872-1879 ◽  
Author(s):  
Bharathwaj Sathyamoorthy ◽  
David M. Parish ◽  
Gaetano T. Montelione ◽  
Rong Xiao ◽  
Thomas Szyperski
Keyword(s):  
Quantum Coherence ◽  
Multiple Quantum ◽  
Nmr Studies ◽  
Biomolecular Nmr ◽  
Heteronuclear Multiple Quantum Coherence ◽  
Coherence Spectroscopy ◽  
Multiple Quantum Coherence

scholarly journals Antimicrobial Peptides Display Strong Synergy with Vancomycin Against Vancomycin-Resistant E. faecium, S. aureus, and Wild-Type E. coli

2020 ◽  
Vol 21 (13) ◽  
pp. 4578 ◽  
Author(s):  
Chih-Lung Wu ◽  
Ju-Yun Hsueh ◽  
Bak-Sau Yip ◽  
Ya-Han Chih ◽  
Kuang-Li Peng ◽  
...  
Keyword(s):  
Antimicrobial Peptides ◽  
Synergistic Effects ◽  
Synergistic Activity ◽  
Wild Type ◽  
Bacterial Strains ◽  
Antibiotic Resistant ◽  
Minimal Inhibitory Concentrations ◽  
E Coli ◽  
Large Size ◽  
Vancomycin Resistant

There is an urgent and imminent need to develop new antimicrobials to fight against antibiotic-resistant bacterial and fungal strains. In this study, a checkerboard method was used to evaluate the synergistic effects of the antimicrobial peptide P-113 and its bulky non-nature amino acid substituted derivatives with vancomycin against vancomycin-resistant Enterococcus faecium, Staphylococcus aureus, and wild-type Escherichia coli. Boron-dipyrro-methene (BODIPY) labeled vancomycin was used to characterize the interactions between the peptides, vancomycin, and bacterial strains. Moreover, neutralization of antibiotic-induced releasing of lipopolysaccharide (LPS) from E. coli by the peptides was obtained. Among these peptides, Bip-P-113 demonstrated the best minimal inhibitory concentrations (MICs), antibiotics synergism, bacterial membrane permeabilization, and supernatant LPS neutralizing activities against the bacteria studied. These results could help in developing antimicrobial peptides that have synergistic activity with large size glycopeptides such as vancomycin in therapeutic applications.

scholarly journals Lectin affinity chromatography and quantitative proteomic analysis reveal that galectin-3 is associated with metastasis in nasopharyngeal carcinoma

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Sathid Aimjongjun ◽  
Onrapak Reamtong ◽  
Tavan Janvilisri
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Affinity Chromatography ◽  
Cell Lines ◽  
Culture Medium ◽  
Lectin Binding ◽  
Therapeutic Interventions ◽  
Migration And Invasion ◽  
Lectin Affinity Chromatography ◽  
Cell Lysates ◽  
Galectin 3

Abstract Nasopharyngeal carcinoma (NPC) is a serious cancer in East and Southeast Asia. Patients are often diagnosed at advanced stages, rendering treatment failure due to high potential of metastasis. This study identified lectin-binding glycoproteins with a potential role in NPC metastasis. Cell lysate and culture medium in highly metastatic 5-8F, and lowly-metastatic 6-10B NPC cell lines were fractionated by ConA- and WGA-affinity chromatography, and subjected to GeLC-MS/MS. A total of 232 and 197 proteins were identified in ConA-enriched fraction of 5-8F and 6-10B cell lysates respectively. In WGA-enriched fraction, 65 and 164 proteins were found in 5-8F and 6-10B cell lysates respectively. Proteins identified in culture medium for both cell lines were 223 and 85 for ConA-enriched fraction, and 94 and 124 for WGA-enriched fraction from 5-8F and 6-10B respectively. Differentially expressed proteins were functionally categorized into cell–cell adhesion, extracellular matrix, glycolysis, protein homeostasis and/or glycosylation enzymes, and lipid metabolism. Interestingly, Galectin-3 (Gal-3) was highly expressed in 5-8F cells but was lowly expressed in 6-10B cells. The Gal-3 knockdown in 5-8F cells, Gal-3 overexpression in 6-10B cells and treatment with Gal-3 inhibitor revealed that Gal-3 was responsible for metastatic phenotypes including adhesion, migration and invasion. So Galectin-3 may serve as a potential target for NPC therapeutic interventions.

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