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The Binding Affinity and Molecular Basis of the Structure-Binding Relationship between Urinary Tamm-Horsfall Glycoprotein and Tumor Necrosis Factor-α
MedLine Citation:
PMID:  23060289     Owner:  NLM     Status:  Publisher    
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.
Cheng-Han Wu; Ko-Jen Li; Sue-Cien Siao; Yu-Hsuan Chen; Tsai-Hung Wu; Chang-Youh Tsai; Chia-Li Yu
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-10-11
Journal Detail:
Title:  Molecules (Basel, Switzerland)     Volume:  17     ISSN:  1420-3049     ISO Abbreviation:  Molecules     Publication Date:  2012  
Date Detail:
Created Date:  2012-10-12     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100964009     Medline TA:  Molecules     Country:  -    
Other Details:
Languages:  ENG     Pagination:  11978-11989     Citation Subset:  -    
Institute of Clinical Medicine, National Taiwan University College of Medicine and National Taiwan University Hospital, No.7 Chung-Shan South Road, Taipei 100, Taiwan.
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