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dc.contributor.authorOk, Kiwon
dc.date.accessioned2021-05-27T14:52:00Z
dc.date.available2021-05-27T14:52:00Z
dc.date.issued2021
dc.identifier.urihttp://hdl.handle.net/10713/15803
dc.descriptionPharmaceutical Sciences
dc.descriptionUniversity of Maryland, Baltimore
dc.descriptionPh.D.
dc.description.abstractZinc (Zn) plays a key role in inflammatory response, including regulating the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway. Among the signaling proteins involved in the NF-κB pathway, many are known zinc finger proteins (ZFs), including Tristetraprolin (TTP). TTP is a non-classical CCCH-type Zinc Finger protein (ZF), that contains two Cys3His zinc binding domains and is a key regulator of the inflammatory response. TTP is a potential target for exogenous gold (Au) and copper (Cu), as well as hydrogen sulfide, an emerging gasotransmitter. To understand how TTP is targeted by other metals, the interactions of TTP were investigated using a combination of bioinorganic chemistry tools including as optical spectroscopy, native electrospray ionization mass spectrometry (ESI-MS), and X-ray absorption study (XAS). The first metal investigated was Cu(I). I discovered that Cu(I) can bind to the tandem ZF construct of TTP (TTP-2D) and disrupt structure and function. This finding indicates a potential relationship between Cu toxicity and metal-regulation of ZFs. The second metal investigated was Au(III). I discovered that the reactivity of TTP-2D with gold complex leads to Au exchange forming a series of Aux-TTP-2D complexes, with reduction of the gold from Au(III) to Au(I). These protein species are then functionally inactive (no RNA binding). When the same experiments were performed with TTP bound to RNA, the Zn-TTP/RNA complex is not disrupted by the Au-complex suggesting a protective role for RNA. To understand how H2S, a signaling molecule, targets Zn-TTP-2D, its reactivity was determined using a combination of cryo-ESI-MS, fluorescence, and electron paramagnetic resonance (EPR) spectroscopies. We found that the H2S oxidizes the cysteine residues of Zn-TTP via a mechanism that involves atmospheric oxygen, a persulfide intermediate and a radical reaction. The results of these biochemical studies of TTP will be presented in the context of TTP’s biological role. In addition, development of a method to follow Zn speciation in inflammatory cells via liquid chromatography connected to inductively coupled plasma (LC-ICP-MS), will be presented. Here, I use THP-1 cells, which are a human monocyte cell line as a model for inflammation, and demonstrate an approach to separate the zinc-proteome.
dc.subjectgold complexesen_US
dc.subjectLC-ICP-MSen_US
dc.subjectmetal ion substitutionen_US
dc.subjectpersulfidationen_US
dc.subjectprotein-RNA interactionsen_US
dc.subjectzinc finger proteinen_US
dc.subject.meshTristetraprolinen_US
dc.titleTargeting Zinc Finger Proteins with Exogenous Metals and Molecules: Lessons Learned from Tristetraprolin, a CCCH type Zinc Fingeren_US
dc.typedissertationen_US
dc.date.updated2021-05-21T13:04:00Z
dc.language.rfc3066en
dc.contributor.advisorMichel, Sarah L. J.
dc.contributor.orcid0000-0002-7724-8860en_US
refterms.dateFOA2021-05-27T14:52:01Z


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