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dc.contributor.authorLee, Cheng-Chungen_US
dc.contributor.authorSu, Yu-Chengen_US
dc.contributor.authorKo, Tzu-Pingen_US
dc.contributor.authorLin, Li-Lingen_US
dc.contributor.authorYang, Chih-Yaen_US
dc.contributor.authorChang, Stanley Shi-Chungen_US
dc.contributor.authorRoffler, Steve R.en_US
dc.contributor.authorWang, Andrew H. -J.en_US
dc.date.accessioned2020-03-02T03:23:23Z-
dc.date.available2020-03-02T03:23:23Z-
dc.date.issued2020-01-07en_US
dc.identifier.issn1021-7770en_US
dc.identifier.urihttp://dx.doi.org/10.1186/s12929-019-0589-7en_US
dc.identifier.urihttp://hdl.handle.net/11536/153702-
dc.description.abstractBackground Polyethylene glycol (PEG) is widely used in industry and medicine. Anti-PEG antibodies have been developed for characterizing PEGylated drugs and other applications. However, the underlying mechanism for specific PEG binding has not been elucidated. Methods The Fab of two cognate anti-PEG antibodies 3.3 and 2B5 were each crystallized in complex with PEG, and their structures were determined by X-ray diffraction. The PEG-Fab interactions in these two crystals were analyzed and compared with those in a PEG-containing crystal of an unrelated anti-hemagglutinin 32D6-Fab. The PEG-binding stoichiometry was examined by using analytical ultracentrifuge (AUC). Results A common PEG-binding mode to 3.3 and 2B5 is seen with an S-shaped core PEG fragment bound to two dyad-related Fab molecules. A nearby satellite binding site may accommodate parts of a longer PEG molecule. The core PEG fragment mainly interacts with the heavy-chain residues D31, W33, L102, Y103 and Y104, making extensive contacts with the aromatic side chains. At the center of each half-circle of the S-shaped PEG, a water molecule makes alternating hydrogen bonds to the ether oxygen atoms, in a similar configuration to that of a crown ether-bound lysine. Each satellite fragment is clamped between two arginine residues, R52 from the heavy chain and R29 from the light chain, and also interacts with several aromatic side chains. In contrast, the non-specifically bound PEG fragments in the 32D6-Fab crystal are located in the elbow region or at lattice contacts. The AUC data suggest that 3.3-Fab exists as a monomer in PEG-free solution but forms a dimer in the presence of PEG-550-MME, which is about the size of the S-shaped core PEG fragment. Conclusions The differing amino acids in 3.3 and 2B5 are not involved in PEG binding but engaged in dimer formation. In particular, the light-chain residue K53 of 2B5-Fab makes significant contacts with the other Fab in a dimer, whereas the corresponding N53 of 3.3-Fab does not. This difference in the protein-protein interaction between two Fab molecules in a dimer may explain the temperature dependence of 2B5 in PEG binding, as well as its inhibition by crown ether.en_US
dc.language.isoen_USen_US
dc.subjectX-ray crystallographyen_US
dc.subjectPEG-fab complexen_US
dc.subjectDimer formationen_US
dc.subjectProtein-protein interactionen_US
dc.subjectCrown etheren_US
dc.titleStructural basis of polyethylene glycol recognition by antibodyen_US
dc.typeArticleen_US
dc.identifier.doi10.1186/s12929-019-0589-7en_US
dc.identifier.journalJOURNAL OF BIOMEDICAL SCIENCEen_US
dc.citation.volume27en_US
dc.citation.issue1en_US
dc.citation.spage0en_US
dc.citation.epage0en_US
dc.contributor.department生物科技學系zh_TW
dc.contributor.departmentDepartment of Biological Science and Technologyen_US
dc.identifier.wosnumberWOS:000512119500001en_US
dc.citation.woscount0en_US
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