Electron spin resonance study of peroxidase activity and kinetics
An electron spin resonance (ESR) assay has been developed for peroxidase activity. The assay measures the formation of the paramagnetic nitroxide Tempol from the oxidation of its hydroxylamine derivative (TOLH) by short-lived radicals produced by peroxidase cycle intermediates, Compounds I and II. U...
Gespeichert in:
| Veröffentlicht in: | Archives of biochemistry and biophysics 1992-11, Vol.299 (1), p.47-56 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 56 |
|---|---|
| container_issue | 1 |
| container_start_page | 47 |
| container_title | Archives of biochemistry and biophysics |
| container_volume | 299 |
| creator | Moore, Katherine L. Moronne, Mario M. Mehlhorn, Rolf J. |
| description | An electron spin resonance (ESR) assay has been developed for peroxidase activity. The assay measures the formation of the paramagnetic nitroxide Tempol from the oxidation of its hydroxylamine derivative (TOLH) by short-lived radicals produced by peroxidase cycle intermediates, Compounds I and II. Using phenol as a peroxidase electron donor, the ESR approach is suitable for measurements of peroxidase activity (⩾0.003 U/ml) and micromolar quantities of H
2O
2 in sample sizes as small as 2 μl. In addition, the ESR method can be used to continuously monitor activity in cell suspensions and other media that are susceptible to optical artifacts. The high membrane permeability of TOLH also makes it possible to estimate peroxidase activity in membrane-enclosed compartments, provided that TOLH oxidation rates can be stimulated with exogenous peroxidase reductants, e.g., phenol. Analysis of TOLH oxidation rates under conditions of low electron donor concentrations and high concentrations of H
2O
2 also shows clear indications of substrate-dependent inhibition and increased catalatic activity. Computer simulations indicate that the results obtained are consistent with the peroxidase reaction scheme proposed by Kohler
et al. (1988,
Arch. Biochem. Biophys. 264, 438–449) modified to correct for a nitroxide dependent stimulation of peroxidase catalytic activity. |
| doi_str_mv | 10.1016/0003-9861(92)90242-O |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_73317568</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>000398619290242O</els_id><sourcerecordid>73317568</sourcerecordid><originalsourceid>FETCH-LOGICAL-c386t-ccf5544f7cc903bf404d09d82b6fc00949200dfa712670b6481bb7118c0265cc3</originalsourceid><addsrcrecordid>eNp90EtLAzEQwPEgitbHN1DYg4geViePZjcXQUp9QKEXPYfsJIHodrcm22K_vVu36M1TDvOfIfwIOadwS4HKOwDguSolvVbsRgETLJ_vkREFJXPgpdgno9_kiByn9A5AqZDskBxSzpmkxYg8TGuHXWybLC1Dk0WX2sY06LLUrewma322dLH9CtYklxnswjp0m8w0NvsIjesCplNy4E2d3NnuPSFvj9PXyXM-mz-9TB5mOfJSdjmiH4-F8AWiAl55AcKCsiWrpEcAJRQDsN4UlMkCKilKWlUFpSUCk2NEfkKuhrvL2H6uXOr0IiR0dW0a166SLjinxViWfSiGEGObUnReL2NYmLjRFPRWTm9Z9JZFK6Z_5PS8X7vY3V9VC2f_lgaqfn65m5uEpvaxZwrpNxOCQ1GKPrsfMtdbrIOLOmFwvagNsZfWtg3__-MbvFqJOQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>73317568</pqid></control><display><type>article</type><title>Electron spin resonance study of peroxidase activity and kinetics</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals Complete</source><creator>Moore, Katherine L. ; Moronne, Mario M. ; Mehlhorn, Rolf J.</creator><creatorcontrib>Moore, Katherine L. ; Moronne, Mario M. ; Mehlhorn, Rolf J.</creatorcontrib><description>An electron spin resonance (ESR) assay has been developed for peroxidase activity. The assay measures the formation of the paramagnetic nitroxide Tempol from the oxidation of its hydroxylamine derivative (TOLH) by short-lived radicals produced by peroxidase cycle intermediates, Compounds I and II. Using phenol as a peroxidase electron donor, the ESR approach is suitable for measurements of peroxidase activity (⩾0.003 U/ml) and micromolar quantities of H
2O
2 in sample sizes as small as 2 μl. In addition, the ESR method can be used to continuously monitor activity in cell suspensions and other media that are susceptible to optical artifacts. The high membrane permeability of TOLH also makes it possible to estimate peroxidase activity in membrane-enclosed compartments, provided that TOLH oxidation rates can be stimulated with exogenous peroxidase reductants, e.g., phenol. Analysis of TOLH oxidation rates under conditions of low electron donor concentrations and high concentrations of H
2O
2 also shows clear indications of substrate-dependent inhibition and increased catalatic activity. Computer simulations indicate that the results obtained are consistent with the peroxidase reaction scheme proposed by Kohler
et al. (1988,
Arch. Biochem. Biophys. 264, 438–449) modified to correct for a nitroxide dependent stimulation of peroxidase catalytic activity.</description><identifier>ISSN: 0003-9861</identifier><identifier>EISSN: 1096-0384</identifier><identifier>DOI: 10.1016/0003-9861(92)90242-O</identifier><identifier>PMID: 1332617</identifier><identifier>CODEN: ABBIA4</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>Analytical, structural and metabolic biochemistry ; Ascorbic Acid - pharmacology ; Biological and medical sciences ; Computer Simulation ; Electron Spin Resonance Spectroscopy - methods ; Enzymes and enzyme inhibitors ; Fundamental and applied biological sciences. Psychology ; Horseradish Peroxidase - antagonists & inhibitors ; Horseradish Peroxidase - metabolism ; Kinetics ; Mathematics ; Models, Theoretical ; Oxidoreductases ; Peroxidases - antagonists & inhibitors ; Peroxidases - metabolism</subject><ispartof>Archives of biochemistry and biophysics, 1992-11, Vol.299 (1), p.47-56</ispartof><rights>1992</rights><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c386t-ccf5544f7cc903bf404d09d82b6fc00949200dfa712670b6481bb7118c0265cc3</citedby><cites>FETCH-LOGICAL-c386t-ccf5544f7cc903bf404d09d82b6fc00949200dfa712670b6481bb7118c0265cc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0003-9861(92)90242-O$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4430784$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1332617$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Moore, Katherine L.</creatorcontrib><creatorcontrib>Moronne, Mario M.</creatorcontrib><creatorcontrib>Mehlhorn, Rolf J.</creatorcontrib><title>Electron spin resonance study of peroxidase activity and kinetics</title><title>Archives of biochemistry and biophysics</title><addtitle>Arch Biochem Biophys</addtitle><description>An electron spin resonance (ESR) assay has been developed for peroxidase activity. The assay measures the formation of the paramagnetic nitroxide Tempol from the oxidation of its hydroxylamine derivative (TOLH) by short-lived radicals produced by peroxidase cycle intermediates, Compounds I and II. Using phenol as a peroxidase electron donor, the ESR approach is suitable for measurements of peroxidase activity (⩾0.003 U/ml) and micromolar quantities of H
2O
2 in sample sizes as small as 2 μl. In addition, the ESR method can be used to continuously monitor activity in cell suspensions and other media that are susceptible to optical artifacts. The high membrane permeability of TOLH also makes it possible to estimate peroxidase activity in membrane-enclosed compartments, provided that TOLH oxidation rates can be stimulated with exogenous peroxidase reductants, e.g., phenol. Analysis of TOLH oxidation rates under conditions of low electron donor concentrations and high concentrations of H
2O
2 also shows clear indications of substrate-dependent inhibition and increased catalatic activity. Computer simulations indicate that the results obtained are consistent with the peroxidase reaction scheme proposed by Kohler
et al. (1988,
Arch. Biochem. Biophys. 264, 438–449) modified to correct for a nitroxide dependent stimulation of peroxidase catalytic activity.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Ascorbic Acid - pharmacology</subject><subject>Biological and medical sciences</subject><subject>Computer Simulation</subject><subject>Electron Spin Resonance Spectroscopy - methods</subject><subject>Enzymes and enzyme inhibitors</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Horseradish Peroxidase - antagonists & inhibitors</subject><subject>Horseradish Peroxidase - metabolism</subject><subject>Kinetics</subject><subject>Mathematics</subject><subject>Models, Theoretical</subject><subject>Oxidoreductases</subject><subject>Peroxidases - antagonists & inhibitors</subject><subject>Peroxidases - metabolism</subject><issn>0003-9861</issn><issn>1096-0384</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp90EtLAzEQwPEgitbHN1DYg4geViePZjcXQUp9QKEXPYfsJIHodrcm22K_vVu36M1TDvOfIfwIOadwS4HKOwDguSolvVbsRgETLJ_vkREFJXPgpdgno9_kiByn9A5AqZDskBxSzpmkxYg8TGuHXWybLC1Dk0WX2sY06LLUrewma322dLH9CtYklxnswjp0m8w0NvsIjesCplNy4E2d3NnuPSFvj9PXyXM-mz-9TB5mOfJSdjmiH4-F8AWiAl55AcKCsiWrpEcAJRQDsN4UlMkCKilKWlUFpSUCk2NEfkKuhrvL2H6uXOr0IiR0dW0a166SLjinxViWfSiGEGObUnReL2NYmLjRFPRWTm9Z9JZFK6Z_5PS8X7vY3V9VC2f_lgaqfn65m5uEpvaxZwrpNxOCQ1GKPrsfMtdbrIOLOmFwvagNsZfWtg3__-MbvFqJOQ</recordid><startdate>19921115</startdate><enddate>19921115</enddate><creator>Moore, Katherine L.</creator><creator>Moronne, Mario M.</creator><creator>Mehlhorn, Rolf J.</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19921115</creationdate><title>Electron spin resonance study of peroxidase activity and kinetics</title><author>Moore, Katherine L. ; Moronne, Mario M. ; Mehlhorn, Rolf J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c386t-ccf5544f7cc903bf404d09d82b6fc00949200dfa712670b6481bb7118c0265cc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Ascorbic Acid - pharmacology</topic><topic>Biological and medical sciences</topic><topic>Computer Simulation</topic><topic>Electron Spin Resonance Spectroscopy - methods</topic><topic>Enzymes and enzyme inhibitors</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Horseradish Peroxidase - antagonists & inhibitors</topic><topic>Horseradish Peroxidase - metabolism</topic><topic>Kinetics</topic><topic>Mathematics</topic><topic>Models, Theoretical</topic><topic>Oxidoreductases</topic><topic>Peroxidases - antagonists & inhibitors</topic><topic>Peroxidases - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Moore, Katherine L.</creatorcontrib><creatorcontrib>Moronne, Mario M.</creatorcontrib><creatorcontrib>Mehlhorn, Rolf J.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Archives of biochemistry and biophysics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Moore, Katherine L.</au><au>Moronne, Mario M.</au><au>Mehlhorn, Rolf J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Electron spin resonance study of peroxidase activity and kinetics</atitle><jtitle>Archives of biochemistry and biophysics</jtitle><addtitle>Arch Biochem Biophys</addtitle><date>1992-11-15</date><risdate>1992</risdate><volume>299</volume><issue>1</issue><spage>47</spage><epage>56</epage><pages>47-56</pages><issn>0003-9861</issn><eissn>1096-0384</eissn><coden>ABBIA4</coden><abstract>An electron spin resonance (ESR) assay has been developed for peroxidase activity. The assay measures the formation of the paramagnetic nitroxide Tempol from the oxidation of its hydroxylamine derivative (TOLH) by short-lived radicals produced by peroxidase cycle intermediates, Compounds I and II. Using phenol as a peroxidase electron donor, the ESR approach is suitable for measurements of peroxidase activity (⩾0.003 U/ml) and micromolar quantities of H
2O
2 in sample sizes as small as 2 μl. In addition, the ESR method can be used to continuously monitor activity in cell suspensions and other media that are susceptible to optical artifacts. The high membrane permeability of TOLH also makes it possible to estimate peroxidase activity in membrane-enclosed compartments, provided that TOLH oxidation rates can be stimulated with exogenous peroxidase reductants, e.g., phenol. Analysis of TOLH oxidation rates under conditions of low electron donor concentrations and high concentrations of H
2O
2 also shows clear indications of substrate-dependent inhibition and increased catalatic activity. Computer simulations indicate that the results obtained are consistent with the peroxidase reaction scheme proposed by Kohler
et al. (1988,
Arch. Biochem. Biophys. 264, 438–449) modified to correct for a nitroxide dependent stimulation of peroxidase catalytic activity.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>1332617</pmid><doi>10.1016/0003-9861(92)90242-O</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-9861 |
| ispartof | Archives of biochemistry and biophysics, 1992-11, Vol.299 (1), p.47-56 |
| issn | 0003-9861 1096-0384 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_73317568 |
| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Analytical, structural and metabolic biochemistry Ascorbic Acid - pharmacology Biological and medical sciences Computer Simulation Electron Spin Resonance Spectroscopy - methods Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology Horseradish Peroxidase - antagonists & inhibitors Horseradish Peroxidase - metabolism Kinetics Mathematics Models, Theoretical Oxidoreductases Peroxidases - antagonists & inhibitors Peroxidases - metabolism |
| title | Electron spin resonance study of peroxidase activity and kinetics |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T05%3A05%3A40IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Electron%20spin%20resonance%20study%20of%20peroxidase%20activity%20and%20kinetics&rft.jtitle=Archives%20of%20biochemistry%20and%20biophysics&rft.au=Moore,%20Katherine%20L.&rft.date=1992-11-15&rft.volume=299&rft.issue=1&rft.spage=47&rft.epage=56&rft.pages=47-56&rft.issn=0003-9861&rft.eissn=1096-0384&rft.coden=ABBIA4&rft_id=info:doi/10.1016/0003-9861(92)90242-O&rft_dat=%3Cproquest_cross%3E73317568%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=73317568&rft_id=info:pmid/1332617&rft_els_id=000398619290242O&rfr_iscdi=true |