https://scholars.lib.ntu.edu.tw/handle/123456789/593989
標題: | The binding affinity and molecular basis of the structure-binding relationship between urinary Tamm-Horsfall glycoprotein and tumor necrosis factor-α | 作者: | CHENG-HAN WU KO-JEN LI Siao, Sue-Cien Chen, Yu-Hsuan Wu, Tsai-Hung Tsai, Chang-Youh CHIA-LI YU |
關鍵字: | Tamm-Horsfall glycoprotein; tumor necrosis factor-alpha; binding affinity; structure-binding relationship; glucosamine-containing mannose; PROTEIN BINDS; LIGHT-CHAINS; UROMODULIN; LECTIN; PHAGOCYTOSIS; ACID; GLYCOSYLATION; LOCALIZATION; SPECIFICITY; CYTOKINES | 公開日期: | 11-十月-2012 | 出版社: | MDPI | 卷: | 17 | 期: | 10 | 起(迄)頁: | 11978 | 來源出版物: | Molecules (Basel, Switzerland) | 摘要: | 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 (K(d)) 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. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/593989 | ISSN: | 1420-3049 | DOI: | 10.3390/molecules171011978 |
顯示於: | 醫學系 |
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