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035 a| (OCoLC)1456021945;

090 a| QR82.B95;

049 a| EREE;

100 1 a| Hart, Kai,e| author.?| UNAUTHORIZED;

245 10 a| Brucella spp. FtrB copper binding thermodynamics and iron oxidase activity /c| by Kai Hart.;

264 1 a| [Greenville, N.C.] :b| [East Carolina University],c| 2024.;

300 a| 1 online resources (82 pages) :b| illustrations (some color);

336 a| textb| txt2| rdacontent;

337 a| computerb| c2| rdamedia;

338 a| online resourceb| cr2| rdacarrier;

347 a| text fileb| Otherc| 3.72 MB2| rda;

538 a| System requirements: Adobe Reader.;

538 a| Mode of access: World Wide Web.;

502 b| M.S.c| East Carolina Universityd| 2024.;

500 a| Presented to the Faculty of the Department of Biology;

500 a| Advisor: Eric Anderson;

500 a| Advisor: Sambuddha Banerjee;

500 a| Title from PDF t.p. (viewed January 15, 2026).;

520 3 a| FtrABCD is a four-component iron uptake system found in several Gram-negative bacteria, including Brucella spp. This four-component transporter consists of two soluble periplasmic subunits (FtrA and FtrB), a putative membrane embedded terminal electron acceptor (FtrD), and a membrane spanning iron permease (FtrC). Based on the evolutionary relationship and sequence homology between FtrC and eukaryotic Ftr1p, this bacterial permease is predicted to be an oxidase dependent Fe2+ transporter. Unlike the eukaryotic Ftr1p permease, which can only function when co-expressed with the multicopper oxidase Fet3p, FtrC does not co-express homologs of any known ferrous oxidase. However, based on its evolutionary relationship with cupredoxins and ferroxidases, periplasmic FtrB is proposed to bind and oxidize Fe2+ during its transport through FtrC. This oxidase property of FtrB can be achieved if it binds a single copper ion using conserved D118 and H121 residues. This non-classical copper ion binding site in FtrB and its ability to form the active enzyme substrate complex (Cu2+ -FtrB-Fe2+) producing the ferrous oxidase function has not been verified experimentally, creating a knowledge gap in the bacterial iron uptake and redox enzyme literature. Data presented in this thesis demonstrates that a) recombinant wild-type Brucella FtrB can form a predominantly [beta]-sheet containing structure; b) the ability of this protein to form this expected native secondary structure and coordinate Cu2+ (Kd = 3.0 ± 1.0 [mu]M in 50 mM bis-tris, 100 mM NaCl, pH 7.3) is dependent on the presence of the conserved residue D118; and c) the recombinant wild-type FtrB can effectively oxidize Fe2+ at pH 7.3 under in vitro conditions. The results described in this thesis are significant as they demonstrate FtrB as a novel ferrous oxidase and open future research opportunities to investigate the importance of this Fe2+ oxidation during iron transport through FtrABCD under in vivo conditions.;

504 a| Includes bibliographical references.;

650 0 a| Brucella.=| ^A1032035;

650 0 a| Brucellosis.=| ^A3503;

650 0 a| Copper proteins.=| ^A12206;

650 0 a| Ironx| Oxidation.=| ^A13130;

653 a| Cupredoxin;

653 a| Copper binding;

653 a| FtrB;

653 a| Ferrous oxidase;

700 1 a| Anderson, Eric,e| degree supervisor.=| ^A696706;

700 1 a| Banerjee, Sambuddha,e| degree supervisor.?| UNAUTHORIZED;

710 2 a| East Carolina University.b| Department of Biology.=| ^A637467;

856 40 z| Access via ScholarShipu| http://hdl.handle.net/10342/13715;

994 a| C0b| ERE;

596 a| 1 4;

998 a| 6477472;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 6477472-1001l| JNETm| JOYNERr| Ys| Yt| JNE3ETDu| 9/16/2024x| ETDz| JERESOURCE;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 6477472-2001l| HSLELECm| HSLr| Ys| Yt| HEETDu| 9/16/2024x| ETDz| HERESOURCE;

Brucella spp. FtrB copper binding thermodynamics and iron oxidase activity / by Kai Hart.

Author/creator	Hart, Kai author.
Other author	Anderson, Eric, degree supervisor.
Other author	Banerjee, Sambuddha, degree supervisor.
Other author	East Carolina University. Department of Biology.
Format	Theses and dissertations
Publication	[Greenville, N.C.] : [East Carolina University], 2024.
Description	1 online resources (82 pages) : illustrations (some color)
Supplemental Content	Access via ScholarShip
Subjects	Brucella. Brucellosis. Copper proteins. Iron--Oxidation.

Summary	FtrABCD is a four-component iron uptake system found in several Gram-negative bacteria, including Brucella spp. This four-component transporter consists of two soluble periplasmic subunits (FtrA and FtrB), a putative membrane embedded terminal electron acceptor (FtrD), and a membrane spanning iron permease (FtrC). Based on the evolutionary relationship and sequence homology between FtrC and eukaryotic Ftr1p, this bacterial permease is predicted to be an oxidase dependent Fe2+ transporter. Unlike the eukaryotic Ftr1p permease, which can only function when co-expressed with the multicopper oxidase Fet3p, FtrC does not co-express homologs of any known ferrous oxidase. However, based on its evolutionary relationship with cupredoxins and ferroxidases, periplasmic FtrB is proposed to bind and oxidize Fe2+ during its transport through FtrC. This oxidase property of FtrB can be achieved if it binds a single copper ion using conserved D118 and H121 residues. This non-classical copper ion binding site in FtrB and its ability to form the active enzyme substrate complex (Cu2+ -FtrB-Fe2+) producing the ferrous oxidase function has not been verified experimentally, creating a knowledge gap in the bacterial iron uptake and redox enzyme literature. Data presented in this thesis demonstrates that a) recombinant wild-type Brucella FtrB can form a predominantly [beta]-sheet containing structure; b) the ability of this protein to form this expected native secondary structure and coordinate Cu2+ (Kd = 3.0 ± 1.0 [mu]M in 50 mM bis-tris, 100 mM NaCl, pH 7.3) is dependent on the presence of the conserved residue D118; and c) the recombinant wild-type FtrB can effectively oxidize Fe2+ at pH 7.3 under in vitro conditions. The results described in this thesis are significant as they demonstrate FtrB as a novel ferrous oxidase and open future research opportunities to investigate the importance of this Fe2+ oxidation during iron transport through FtrABCD under in vivo conditions.
General note	Presented to the Faculty of the Department of Biology
General note	Advisor: Eric Anderson
General note	Advisor: Sambuddha Banerjee
General note	Title from PDF t.p. (viewed January 15, 2026).
Dissertation note	M.S. East Carolina University 2024.
Bibliography note	Includes bibliographical references.
Technical details	System requirements: Adobe Reader.
Technical details	Mode of access: World Wide Web.

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Brucella spp. FtrB copper binding thermodynamics and iron oxidase activity / by Kai Hart.

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