Please use this identifier to cite or link to this item:
Full metadata record
DC FieldValueLanguage
dc.contributor.authorToh, Yew Kwangen_US
dc.contributor.authorShin, Joonen_US
dc.contributor.authorBalakrishna, Asha Manikkothen_US
dc.contributor.authorKamariah, Neelagandanen_US
dc.contributor.authorGrüber, Ardinaen_US
dc.contributor.authorEisenhaber, Franken_US
dc.contributor.authorEisenhaber, Birgiten_US
dc.contributor.authorGrüber, Gerharden_US
dc.identifier.citationToh, Y. K., Shin, J., Balakrishna, A. M., Kamariah, N., Grüber, A., Eisenhaber, F., . . . Grüber, G. (2019). Effect of the additional cysteine 503 of vancomycin-resistant Enterococcus faecalis (V583) alkylhydroperoxide reductase subunit F (AhpF) and the mechanism of AhpF and subunit C assembling. Free Radical Biology and Medicine, 138, 10–22. doi:10.1016/j.freeradbiomed.2019.04.036en_US
dc.description.abstractThe vancomycin-resistant Enterococcus faecalis alkyl hydroperoxide reductase complex (AhpR) with its subunits AhpC (EfAhpC) and AhpF (EfAhpF) is of paramount importance to restore redox homeostasis. Therefore, knowledge about this defense system is essential to understand its antibiotic-resistance and survival in hosts. Recently, we described the crystallographic structures of EfAhpC, the two-fold thioredoxin-like domain of EfAhpF, the novel phenomenon of swapping of the catalytic domains of EfAhpF as well as the unique linker length, connecting the catalytically active N-and C-terminal domains of EfAhpF. Here, using mutagenesis and enzymatic studies, we reveal the effect of an additional third cysteine (C503) in EfAhpF, which might optimize the functional adaptation of the E. faecalis enzyme under various physiological conditions. The crystal structure and solution NMR data of the engineered C503A mutant of the thioredoxin-like domain of EfAhpF were used to describe alterations in the environment of the additional cysteine residue during modulation of the redox-state. To glean insight into the epitope and mechanism of EfAhpF and -AhpC interaction as well as the electron transfer from the thioredoxin-like domain of EfAhpF to AhpC, NMR-titration experiments were performed, showing a coordinated disappearance of peaks in the thioredoxin-like domain of EfAhpF in the presence of full length EfAhpC, and indicating a stable EfAhpF-AhpC-complex. Combined with docking studies, the interacting residues of EfAhpF were identified and a mechanism of electron transfer of the EfAhpF donor to the electron acceptor EfAhpC is described.en_US
dc.description.sponsorshipMinistry of Education (MOE)en_US
dc.relation.ispartofFree radical biology & medicineen_US
dc.rights© 2019 Elsevier Inc. All rights reserved. This paper was published in Free radical biology & medicine and is made available with permission of Elsevier Inc.en_US
dc.subjectScience::Biological sciencesen_US
dc.titleEffect of the additional cysteine 503 of vancomycin-resistant Enterococcus faecalis (V583) alkylhydroperoxide reductase subunit F (AhpF) and the mechanism of AhpF and subunit C assemblingen_US
dc.typeJournal Articleen
dc.contributor.schoolSchool of Computer Science and Engineeringen_US
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.contributor.organizationAgency for Science, Technology and Researchen_US
dc.description.versionAccepted versionen_US
dc.subject.keywordsReactive Oxygen Speciesen_US
dc.subject.keywordsOxidative Stressen_US
dc.description.acknowledgementThis research was supported by a Singapore Ministry of Education Academic Research Fund Tier 1 (RG140/16) to G.G. (M4080811.080). We thank Dr. S. S. M. Malathy for the art work of Fig. 1A.en_US
item.fulltextWith Fulltext-
Appears in Collections:SBS Journal Articles

Citations 50

Updated on Sep 23, 2023

Web of ScienceTM
Citations 50

Updated on Sep 25, 2023

Page view(s)

Updated on Sep 23, 2023

Download(s) 50

Updated on Sep 23, 2023

Google ScholarTM




Items in DR-NTU are protected by copyright, with all rights reserved, unless otherwise indicated.