{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/70533"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/70533","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Glutamine Phosphoribosyl Pyrophosphate Amidotransferase From Bacillus Subtilis: Iron-Sulfur Biochemistry and Enzymology","abstract":"The iron-sulfur prosthetic group of glutamine PRPP amidotransferase was found to be a {4Fe-4S} cluster. In the native enzyme, the cluster exists in the diamagnetic +2 redox state. Treatment of the cluster with oxidants resulted either in no reaction or in oxidative dissolution of the cluster, i.e., a +3 state was not detected. The cluster was reduced poorly with sodium dithionite, but was reduced readily by photoreduction with 5-deazaflavin. Photoreduction of the native enzyme resulted in formation of the +1 redox state of the cluster. The reduced enzyme was EPR silent. Examination of the reduced enzyme by Mossbauer spectroscopy indicated the presence of multiple unpaired electrons (S (GREATERTHEQ) 3/2). The E(,m) of the +1/+2 couple was determined to be (LESSTHEQ) -620 mV. The reduced enzyme was found to be active in the following assays: glutamine PRPP amidotransferase, glutaminase and PRPP hydrolase; these results demonstrated that the cluster does not have a redox function during catalysis of amide transfer by the enzyme.","abstract_html":"The iron-sulfur prosthetic group of glutamine PRPP amidotransferase was found to be a {4Fe-4S} cluster. In the native enzyme, the cluster exists in the diamagnetic +2 redox state. Treatment of the cluster with oxidants resulted either in no reaction or in oxidative dissolution of the cluster, i.e., a +3 state was not detected. The cluster was reduced poorly with sodium dithionite, but was reduced readily by photoreduction with 5-deazaflavin. Photoreduction of the native enzyme resulted in formation of the +1 redox state of the cluster. The reduced enzyme was EPR silent. Examination of the reduced enzyme by Mossbauer spectroscopy indicated the presence of multiple unpaired electrons (S (GREATERTHEQ) 3/2). The E(,m) of the +1/+2 couple was determined to be (LESSTHEQ) -620 mV. The reduced enzyme was found to be active in the following assays: glutamine PRPP amidotransferase, glutaminase and PRPP hydrolase; these results demonstrated that the cluster does not have a redox function during catalysis of amide transfer by the enzyme.","abstract_has_math":false,"creators":["Vollmer, Steven John"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Biochemistry","degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-12-15T23:43:47Z","date_published":"2014-12-15T23:43:47Z","updated_at":"2026-07-22T22:26:03Z","subjects":["Chemistry, Biochemistry"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["(UMI)AAI8410066"],"render_values":[{"text":"(UMI)AAI8410066","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/70533","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Vollmer, Steven John"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2014-12-15T23:43:47Z","10000-01-01","1983"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biochemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Chemistry, Biochemistry"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/70533","(UMI)AAI8410066"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The iron-sulfur prosthetic group of glutamine PRPP amidotransferase was found to be a {4Fe-4S} cluster. In the native enzyme, the cluster exists in the diamagnetic +2 redox state. Treatment of the cluster with oxidants resulted either in no reaction or in oxidative dissolution of the cluster, i.e., a +3 state was not detected. The cluster was reduced poorly with sodium dithionite, but was reduced readily by photoreduction with 5-deazaflavin. Photoreduction of the native enzyme resulted in formation of the +1 redox state of the cluster. The reduced enzyme was EPR silent. Examination of the reduced enzyme by Mossbauer spectroscopy indicated the presence of multiple unpaired electrons (S (GREATERTHEQ) 3/2). The E(,m) of the +1/+2 couple was determined to be (LESSTHEQ) -620 mV. The reduced enzyme was found to be active in the following assays: glutamine PRPP amidotransferase, glutaminase and PRPP hydrolase; these results demonstrated that the cluster does not have a redox function during catalysis of amide transfer by the enzyme.","The glutamine utilizing site of the enzyme was examined through the use of the affinity ligand 6-diazo-5-oxo-L-norleucine. The thiolate of Cys-1 was identified as the enzyme nucleophile that hydrolyzes glutamine during amide transfer. The complete amino acid sequence of the enzyme is available, and the ligands to the iron-sulfur clusters have been tentatively identified: Cys-382, Cys-434, Cys-437 and Cys-440.","Made available in DSpace on 2014-12-15T23:43:47Z (GMT). No. of bitstreams: 1 8410066.pdf: 8836683 bytes, checksum: 8b7b406dcafd62c497f715821d4ffb30 (MD5) Previous issue date: 1983","Embargo set by: Seth Robbins for item 70699 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","279 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1983."]},{"key":"dc:title","label":"Title","values":["Glutamine Phosphoribosyl Pyrophosphate Amidotransferase From Bacillus Subtilis: Iron-Sulfur Biochemistry and Enzymology"]}]}],"canonical_facts":{"dc:creator":["Vollmer, Steven John"],"dc:date":["2014-12-15T23:43:47Z","10000-01-01","1983"],"dc:description":["The iron-sulfur prosthetic group of glutamine PRPP amidotransferase was found to be a {4Fe-4S} cluster. In the native enzyme, the cluster exists in the diamagnetic +2 redox state. Treatment of the cluster with oxidants resulted either in no reaction or in oxidative dissolution of the cluster, i.e., a +3 state was not detected. The cluster was reduced poorly with sodium dithionite, but was reduced readily by photoreduction with 5-deazaflavin. Photoreduction of the native enzyme resulted in formation of the +1 redox state of the cluster. The reduced enzyme was EPR silent. Examination of the reduced enzyme by Mossbauer spectroscopy indicated the presence of multiple unpaired electrons (S (GREATERTHEQ) 3/2). The E(,m) of the +1/+2 couple was determined to be (LESSTHEQ) -620 mV. The reduced enzyme was found to be active in the following assays: glutamine PRPP amidotransferase, glutaminase and PRPP hydrolase; these results demonstrated that the cluster does not have a redox function during catalysis of amide transfer by the enzyme.","The glutamine utilizing site of the enzyme was examined through the use of the affinity ligand 6-diazo-5-oxo-L-norleucine. The thiolate of Cys-1 was identified as the enzyme nucleophile that hydrolyzes glutamine during amide transfer. The complete amino acid sequence of the enzyme is available, and the ligands to the iron-sulfur clusters have been tentatively identified: Cys-382, Cys-434, Cys-437 and Cys-440.","Made available in DSpace on 2014-12-15T23:43:47Z (GMT). No. of bitstreams: 1 8410066.pdf: 8836683 bytes, checksum: 8b7b406dcafd62c497f715821d4ffb30 (MD5) Previous issue date: 1983","Embargo set by: Seth Robbins for item 70699 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","279 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1983."],"dc:identifier":["http://hdl.handle.net/2142/70533","(UMI)AAI8410066"],"dc:subject":["Chemistry, Biochemistry"],"dc:title":["Glutamine Phosphoribosyl Pyrophosphate Amidotransferase From Bacillus Subtilis: Iron-Sulfur Biochemistry and Enzymology"],"dc:type":["text"],"thesis:degree_discipline":["Biochemistry"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:26:03Z"}