The Binding Affinity and Molecular Basis of the Structure-Binding Relationship between Urinary Tamm-Horsfall Glycoprotein and Tumor Necrosis Factor-α
Open Access
- 10 October 2012
- Vol. 17 (10), 11978-11989
- https://doi.org/10.3390/molecules171011978
Abstract
In a previous study we noted significant THP binding to TNF-α, but did not explore the molecular basis of the structure-binding relationship. In this study, we used lectin-binding ELISA to assess the carbohydrate compositions of THP, BSA, IgG, TNF-α, and IFN-g. We identified β(1,4)-N-acetylglucosamine oligomers (GlcNAc) and GlcNAc/branched mannose in BSA, IgG, TNF-α, and THP, but not in IFN-g. These carbohydrate moieties mediated binding with THP. Small amounts of Siaα(2,3)Gal/ GalNAc, Sia(2,6)Gal/GalNAc, and mannose residues were also present in THP and TNF-α. Binding affinity (Kd) between THP and TNF-α by Scatchard plot analysis was 1.4–1.7 × 10−6 M, lower than antigen-antibody or ligand-receptor binding affinities. To elucidate the structure-binding relationship of THP-TNF-α, THP was digested with neuraminidase, β-galactosidase, O-sialoglycoprotein endopeptidase, carboxypeptidase Y, or proteinase K. β-galactosidase increased binding capacity of THP for TNF-α. Monosaccharide inhibition suggested that α-methyl-D-mannoside, GlcNAc, and GalNAc, but not sialic acid, suppress THP-TNF-α binding as detected by ELISA. We conclude that sugar-lectin and sugar-protein interactions between cognate sites in THP and TNF-α mediate their binding.Keywords
This publication has 38 references indexed in Scilit:
- Systematic Review and Meta-Analysis of the Efficacy and Safety of Existing TNF Blocking Agents in Treatment of Rheumatoid ArthritisPLOS ONE, 2012
- Tamm-Horsfall Glycoprotein Enhances PMN Phagocytosis by Binding to Cell Surface-Expressed Lactoferrin and Cathepsin G That Activates MAP Kinase PathwayMolecules, 2011
- The efficacy of anti-TNF in rheumatoid arthritis, a comparison between randomised controlled trials and clinical practiceAnnals Of The Rheumatic Diseases, 2007
- Potential new targets in arthritis therapy: interleukin (IL)-17 and its relation to tumour necrosis factor and IL-1 in experimental arthritisAnnals Of The Rheumatic Diseases, 2006
- Binding of Tamm‐Horsfall protein to complement 1q measured by ELISA and resonant mirror biosensor techniques under various ionic‐strength conditionsImmunology & Cell Biology, 2000
- Interleukin 8, Neutrophil-Activating Peptide-2 and GRO-α Bind to and Elicit Cell Activation via Specific and Different Amino Acid Residues of CXCR2Cytokine, 2000
- Localization of a single binding site for immunoglobulin light chains on human Tamm-Horsfall glycoprotein.JCI Insight, 1997
- Tamm-Horsfall glycoprotein binds IgG with high affinityKidney International, 1993
- Tamm Horsfall protein binds to a single class of carbohydrate specific receptors on human neutrophilsKidney International, 1993
- Native cytokines do not bind to uromodulin (Tamm‐Horsfall glycoprotein)FEBS Letters, 1988