Molecules 2012, 17(10), 11978-11989; doi:10.3390/molecules171011978
Article

The Binding Affinity and Molecular Basis of the Structure-Binding Relationship between Urinary Tamm-Horsfall Glycoprotein and Tumor Necrosis Factor-α

1,†, 2,†, 3, 3, 4, 5 and 3,* email
Received: 13 July 2012; in revised form: 26 September 2012 / Accepted: 8 October 2012 / Published: 11 October 2012
(This article belongs to the Special Issue Advances in Carbohydrate Chemistry)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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: Tamm-Horsfall glycoprotein; tumor necrosis factor-α; binding affinity; structure-binding relationship; glucosamine-containing mannose
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MDPI and ACS Style

Wu, C.-H.; Li, K.-J.; Siao, S.-C.; Chen, Y.-H.; Wu, T.-H.; Tsai, C.-Y.; Yu, C.-L. The Binding Affinity and Molecular Basis of the Structure-Binding Relationship between Urinary Tamm-Horsfall Glycoprotein and Tumor Necrosis Factor-α. Molecules 2012, 17, 11978-11989.

AMA Style

Wu C-H, Li K-J, Siao S-C, Chen Y-H, Wu T-H, Tsai C-Y, Yu C-L. The Binding Affinity and Molecular Basis of the Structure-Binding Relationship between Urinary Tamm-Horsfall Glycoprotein and Tumor Necrosis Factor-α. Molecules. 2012; 17(10):11978-11989.

Chicago/Turabian Style

Wu, Cheng-Han; Li, Ko-Jen; Siao, Sue-Cien; Chen, Yu-Hsuan; Wu, Tsai-Hung; Tsai, Chang-Youh; Yu, Chia-Li. 2012. "The Binding Affinity and Molecular Basis of the Structure-Binding Relationship between Urinary Tamm-Horsfall Glycoprotein and Tumor Necrosis Factor-α." Molecules 17, no. 10: 11978-11989.


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