{"id":{"repo_id":"usf","oai_identifier":"oai:digitalcommons.usf.edu:etd-1072"},"canonical_url":"https://search.dev.ndltd.org/etd/usf/oai:digitalcommons.usf.edu:etd-1072","repository":{"repo_id":"usf","name":"University of South Florida","base_url":"https://digitalcommons.usf.edu/do/oai/"},"display":{"title":"Two Methodologies in Pursuit of the Elucidation of Copper (II)—Centered Bioinorganic Chemistry","abstract":"Copper is a widely distributed transition metal in the earth's crust and has been adopted in a variety of biological systems. In many ways the biochemical usefulness of copper stems from its positive redox potential. This positive redox potential allows copper to assist in the movement of electrons. Copper ions can be found in natural systems as either CuI, CuII or CuIII in part due to this redox potential. While CuII -centered biochemistry has been studied for years, mechanistic details in certain CuII -centered redox reactions remain unresolved. This study presents two methodologies for studying natural systems with known CuII -centered redox capabilities in order to better elucidate the mechanistic intricacies of Copper ion chemistry. The first method explored involves the promiscuous enzyme Streptomyces griseus aminopeptidase (SgAP) which although known primarily as a peptidase has been shown to oxidize catechol under near physiological conditions in vitro when its native ZnII ions are replaced by CuII ions. Protein engineering techniques were utilized toward expression a functional recombinant enzyme in wild type and mutant forms. The goal was to utilize Site directed mutagenesis of residues in the active site to determine which residues are involved in both the hydrolysis and the oxidative activities of SgAP. The second methodology explored was the use of the N-terminus of Histatin-5, a naturally occurring peptide that is known to form complexes with CuII, as a model system to study CuII -centered oxidation chemistry. Metal-Peptide complexes are much more simplified model systems which use the same building blocks as proteins, but reduce the structure to the minimal functional unit necessary for activity. This in turn, simplifies the study of their catalytic chemistry as influences outside of the active region are greatly reduced. Furthermore, chemical synthesis of short peptides is easily performed and inexpensive in comparison to protein engineering, thus enabling further exploration, if deemed necessary, to be a feasible and economically viable possibility.","abstract_html":"Copper is a widely distributed transition metal in the earth's crust and has been adopted in a variety of biological systems. In many ways the biochemical usefulness of copper stems from its positive redox potential. This positive redox potential allows copper to assist in the movement of electrons. Copper ions can be found in natural systems as either CuI, CuII or CuIII in part due to this redox potential. While CuII -centered biochemistry has been studied for years, mechanistic details in certain CuII -centered redox reactions remain unresolved. This study presents two methodologies for studying natural systems with known CuII -centered redox capabilities in order to better elucidate the mechanistic intricacies of Copper ion chemistry. The first method explored involves the promiscuous enzyme Streptomyces griseus aminopeptidase (SgAP) which although known primarily as a peptidase has been shown to oxidize catechol under near physiological conditions in vitro when its native ZnII ions are replaced by CuII ions. Protein engineering techniques were utilized toward expression a functional recombinant enzyme in wild type and mutant forms. The goal was to utilize Site directed mutagenesis of residues in the active site to determine which residues are involved in both the hydrolysis and the oxidative activities of SgAP. The second methodology explored was the use of the N-terminus of Histatin-5, a naturally occurring peptide that is known to form complexes with CuII, as a model system to study CuII -centered oxidation chemistry. Metal-Peptide complexes are much more simplified model systems which use the same building blocks as proteins, but reduce the structure to the minimal functional unit necessary for activity. This in turn, simplifies the study of their catalytic chemistry as influences outside of the active region are greatly reduced. Furthermore, chemical synthesis of short peptides is easily performed and inexpensive in comparison to protein engineering, thus enabling further exploration, if deemed necessary, to be a feasible and economically viable possibility.","abstract_has_math":false,"creators":["Wagner, William John"],"institution":"Digital Commons @ University of South Florida","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2009,"date_issued":"2009-03-30T07:00:00Z","date_published":"2009-03-30T07:00:00Z","updated_at":"2026-07-24T05:42:29Z","subjects":["Aminopeptidase","Streptomyces","Copper","Moonlighting","Histatin","American Studies","Arts and Humanities"],"languages":[],"rights":["default"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.usf.edu/etd/73","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Wagner, William John"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2009-03-30T07:00:00Z"]},{"key":"dc:publisher","label":"Institution","values":["Digital Commons @ University of South Florida"]},{"key":"dc:type","label":"Dc Type","values":["thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Aminopeptidase","Streptomyces","Copper","Moonlighting","Histatin","American Studies","Arts and Humanities"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:rights","label":"Dc Rights","values":["default"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.usf.edu/etd/73","https://digitalcommons.usf.edu/context/etd/article/1072/viewcontent/etd__73.pdf"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Copper is a widely distributed transition metal in the earth's crust and has been adopted in a variety of biological systems. In many ways the biochemical usefulness of copper stems from its positive redox potential. This positive redox potential allows copper to assist in the movement of electrons. Copper ions can be found in natural systems as either CuI, CuII or CuIII in part due to this redox potential. While CuII -centered biochemistry has been studied for years, mechanistic details in certain CuII -centered redox reactions remain unresolved. This study presents two methodologies for studying natural systems with known CuII -centered redox capabilities in order to better elucidate the mechanistic intricacies of Copper ion chemistry. The first method explored involves the promiscuous enzyme Streptomyces griseus aminopeptidase (SgAP) which although known primarily as a peptidase has been shown to oxidize catechol under near physiological conditions in vitro when its native ZnII ions are replaced by CuII ions. Protein engineering techniques were utilized toward expression a functional recombinant enzyme in wild type and mutant forms. The goal was to utilize Site directed mutagenesis of residues in the active site to determine which residues are involved in both the hydrolysis and the oxidative activities of SgAP. The second methodology explored was the use of the N-terminus of Histatin-5, a naturally occurring peptide that is known to form complexes with CuII, as a model system to study CuII -centered oxidation chemistry. Metal-Peptide complexes are much more simplified model systems which use the same building blocks as proteins, but reduce the structure to the minimal functional unit necessary for activity. This in turn, simplifies the study of their catalytic chemistry as influences outside of the active region are greatly reduced. Furthermore, chemical synthesis of short peptides is easily performed and inexpensive in comparison to protein engineering, thus enabling further exploration, if deemed necessary, to be a feasible and economically viable possibility."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:source","label":"Dc Source","values":["USF Tampa Graduate Theses and Dissertations"]},{"key":"dc:title","label":"Title","values":["Two Methodologies in Pursuit of the Elucidation of Copper (II)—Centered Bioinorganic Chemistry"]}]}],"canonical_facts":{"dc:creator":["Wagner, William John"],"dc:date":["2009-03-30T07:00:00Z"],"dc:description":["Copper is a widely distributed transition metal in the earth's crust and has been adopted in a variety of biological systems. In many ways the biochemical usefulness of copper stems from its positive redox potential. This positive redox potential allows copper to assist in the movement of electrons. Copper ions can be found in natural systems as either CuI, CuII or CuIII in part due to this redox potential. While CuII -centered biochemistry has been studied for years, mechanistic details in certain CuII -centered redox reactions remain unresolved. This study presents two methodologies for studying natural systems with known CuII -centered redox capabilities in order to better elucidate the mechanistic intricacies of Copper ion chemistry. The first method explored involves the promiscuous enzyme Streptomyces griseus aminopeptidase (SgAP) which although known primarily as a peptidase has been shown to oxidize catechol under near physiological conditions in vitro when its native ZnII ions are replaced by CuII ions. Protein engineering techniques were utilized toward expression a functional recombinant enzyme in wild type and mutant forms. The goal was to utilize Site directed mutagenesis of residues in the active site to determine which residues are involved in both the hydrolysis and the oxidative activities of SgAP. The second methodology explored was the use of the N-terminus of Histatin-5, a naturally occurring peptide that is known to form complexes with CuII, as a model system to study CuII -centered oxidation chemistry. Metal-Peptide complexes are much more simplified model systems which use the same building blocks as proteins, but reduce the structure to the minimal functional unit necessary for activity. This in turn, simplifies the study of their catalytic chemistry as influences outside of the active region are greatly reduced. Furthermore, chemical synthesis of short peptides is easily performed and inexpensive in comparison to protein engineering, thus enabling further exploration, if deemed necessary, to be a feasible and economically viable possibility."],"dc:format":["application/pdf"],"dc:identifier":["https://digitalcommons.usf.edu/etd/73","https://digitalcommons.usf.edu/context/etd/article/1072/viewcontent/etd__73.pdf"],"dc:publisher":["Digital Commons @ University of South Florida"],"dc:rights":["default"],"dc:source":["USF Tampa Graduate Theses and Dissertations"],"dc:subject":["Aminopeptidase","Streptomyces","Copper","Moonlighting","Histatin","American Studies","Arts and Humanities"],"dc:title":["Two Methodologies in Pursuit of the Elucidation of Copper (II)—Centered Bioinorganic Chemistry"],"dc:type":["thesis"]},"updated_at":"2026-07-24T05:42:29Z"}