Reduction of Clofazimine by Mycobacterial Type 2 NADH:Quinone Oxidoreductase

The mechanism of action of clofazimine (CFZ), an antimycobacterial drug with a long history, is not well understood. The present study describes a redox cycling pathway that involves the enzymatic reduction of CFZ by NDH-2, the primary respiratory chain NADH:quinone oxidoreductase of mycobacteria an...

Full description

Saved in:

Bibliographic Details
Published in:	The Journal of biological chemistry 2011-03, Vol.286 (12), p.10276-10287
Main Authors:	Yano, Takahiro, Kassovska-Bratinova, Sacha, Teh, J. Shin, Winkler, Jeffrey, Sullivan, Kevin, Isaacs, Andre, Schechter, Norman M., Rubin, Harvey
Format:	Article
Language:	eng
Subjects:	Amyloid Blood Brain RNA Binding Protein RNA Folding Translation Control
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

recordid	cdi_crossref_primary_10_1074_jbc_M110_200501
title	Reduction of Clofazimine by Mycobacterial Type 2 NADH:Quinone Oxidoreductase
format	Article
creator	Yano, Takahiro Kassovska-Bratinova, Sacha Teh, J. Shin Winkler, Jeffrey Sullivan, Kevin Isaacs, Andre Schechter, Norman M. Rubin, Harvey
subjects	Amyloid Blood Brain RNA Binding Protein RNA Folding Translation Control
ispartof	The Journal of biological chemistry, 2011-03, Vol.286 (12), p.10276-10287
description	The mechanism of action of clofazimine (CFZ), an antimycobacterial drug with a long history, is not well understood. The present study describes a redox cycling pathway that involves the enzymatic reduction of CFZ by NDH-2, the primary respiratory chain NADH:quinone oxidoreductase of mycobacteria and nonenzymatic oxidation of reduced CFZ by O2 yielding CFZ and reactive oxygen species (ROS). This pathway was demonstrated using isolated membranes and purified recombinant NDH-2. The reduction and oxidation of CFZ was measured spectrally, and the production of ROS was measured using a coupled assay system with Amplex Red. Supporting the ROS-based killing mechanism, bacteria grown in the presence of antioxidants are more resistant to CFZ. CFZ-mediated increase in NADH oxidation and ROS production were not observed in membranes from three different Gram-negative bacteria but was observed in Staphylococcus aureus and Saccharomyces cerevisiae, which is consistent with the known antimicrobial specificity of CFZ. A more soluble analog of CFZ, KS6, was synthesized and was shown to have the same activities as CFZ. These studies describe a pathway for a continuous and high rate of reactive oxygen species production in Mycobacterium smegmatis treated with CFZ and a CFZ analog as well as evidence that cell death produced by these agents are related to the production of these radical species.
language	eng
source	BACON - Elsevier - GLOBAL_SCIENCEDIRECT-OPENACCESS; Open Access: PubMed Central; ScienceDirect (Online service)
identifier	ISSN: 0021-9258
fulltext	fulltext
issn	0021-9258 1083-351X
url	http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-05-27T07%3A51%3A36IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-elsevier_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Reduction%20of%20Clofazimine%20by%20Mycobacterial%20Type%202%20NADH:Quinone%20Oxidoreductase&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Yano,%20Takahiro&rft.date=2011-03&rft.volume=286&rft.issue=12&rft.spage=10276&rft.epage=10287&rft.pages=10276-10287&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.M110.200501&rft_dat=%3Celsevier_cross%3ES0021925820538544%3C/elsevier_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c2431-5698a12eb14921f5a625bd8617db855552ddebe56b16c538173a0110fa235d3a3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_id=info:pmid/
container_title	The Journal of biological chemistry
container_volume	286
container_issue	12
container_start_page	10276
container_end_page	10287
fullrecord	<record><control><sourceid>elsevier_cross</sourceid><recordid>TN_cdi_crossref_primary_10_1074_jbc_M110_200501</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0021925820538544</els_id><sourcerecordid>S0021925820538544</sourcerecordid><originalsourceid>FETCH-LOGICAL-c2431-5698a12eb14921f5a625bd8617db855552ddebe56b16c538173a0110fa235d3a3</originalsourceid><addsrcrecordid>eNp1kM1LAzEQxYMoWKtnr_kHts0km-2ut1I_KmwtSgVvIR-zkNJuSrYV17_e1Hr1XYaBeY95P0JugY2ATfLx2tjRAtLGGZMMzsgAWCkyIeHjnAwY45BVXJaX5Krr1iwpr2BA6jd0B7v3oaWhobNNaPS33_oWqenporfBaLvH6PWGrvodUk5fpvfzu9eDb0M6Wn55F-JvhO7wmlw0etPhzd8ckvfHh9VsntXLp-fZtM4szwVksqhKDRwN5BWHRuqCS-PKAibOlDKJO4cGZWGgsFKUMBGapWaN5kI6ocWQjE-5Noaui9ioXfRbHXsFTB1hqARDHWGoE4zkqE4OTG99eoyqsx5bi85HtHvlgv_X-wMRkWS5</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><isCDI>true</isCDI><recordtype>article</recordtype></control><display><type>article</type><title>Reduction of Clofazimine by Mycobacterial Type 2 NADH:Quinone Oxidoreductase</title><source>BACON - Elsevier - GLOBAL_SCIENCEDIRECT-OPENACCESS</source><source>Open Access: PubMed Central</source><source>ScienceDirect (Online service)</source><creator>Yano, Takahiro ; Kassovska-Bratinova, Sacha ; Teh, J. Shin ; Winkler, Jeffrey ; Sullivan, Kevin ; Isaacs, Andre ; Schechter, Norman M. ; Rubin, Harvey</creator><creatorcontrib>Yano, Takahiro ; Kassovska-Bratinova, Sacha ; Teh, J. Shin ; Winkler, Jeffrey ; Sullivan, Kevin ; Isaacs, Andre ; Schechter, Norman M. ; Rubin, Harvey</creatorcontrib><description>The mechanism of action of clofazimine (CFZ), an antimycobacterial drug with a long history, is not well understood. The present study describes a redox cycling pathway that involves the enzymatic reduction of CFZ by NDH-2, the primary respiratory chain NADH:quinone oxidoreductase of mycobacteria and nonenzymatic oxidation of reduced CFZ by O2 yielding CFZ and reactive oxygen species (ROS). This pathway was demonstrated using isolated membranes and purified recombinant NDH-2. The reduction and oxidation of CFZ was measured spectrally, and the production of ROS was measured using a coupled assay system with Amplex Red. Supporting the ROS-based killing mechanism, bacteria grown in the presence of antioxidants are more resistant to CFZ. CFZ-mediated increase in NADH oxidation and ROS production were not observed in membranes from three different Gram-negative bacteria but was observed in Staphylococcus aureus and Saccharomyces cerevisiae, which is consistent with the known antimicrobial specificity of CFZ. A more soluble analog of CFZ, KS6, was synthesized and was shown to have the same activities as CFZ. These studies describe a pathway for a continuous and high rate of reactive oxygen species production in Mycobacterium smegmatis treated with CFZ and a CFZ analog as well as evidence that cell death produced by these agents are related to the production of these radical species.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M110.200501</identifier><language>eng</language><publisher>Elsevier Inc</publisher><subject>Amyloid ; Blood ; Brain ; RNA Binding Protein ; RNA Folding ; Translation Control</subject><ispartof>The Journal of biological chemistry, 2011-03, Vol.286 (12), p.10276-10287</ispartof><rights>2011 © 2011 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c2431-5698a12eb14921f5a625bd8617db855552ddebe56b16c538173a0110fa235d3a3</citedby><cites>FETCH-LOGICAL-c2431-5698a12eb14921f5a625bd8617db855552ddebe56b16c538173a0110fa235d3a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0021925820538544$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>315,787,791,3569,27985,27986,46162</link.rule.ids></links><search><creatorcontrib>Yano, Takahiro</creatorcontrib><creatorcontrib>Kassovska-Bratinova, Sacha</creatorcontrib><creatorcontrib>Teh, J. Shin</creatorcontrib><creatorcontrib>Winkler, Jeffrey</creatorcontrib><creatorcontrib>Sullivan, Kevin</creatorcontrib><creatorcontrib>Isaacs, Andre</creatorcontrib><creatorcontrib>Schechter, Norman M.</creatorcontrib><creatorcontrib>Rubin, Harvey</creatorcontrib><title>Reduction of Clofazimine by Mycobacterial Type 2 NADH:Quinone Oxidoreductase</title><title>The Journal of biological chemistry</title><description>The mechanism of action of clofazimine (CFZ), an antimycobacterial drug with a long history, is not well understood. The present study describes a redox cycling pathway that involves the enzymatic reduction of CFZ by NDH-2, the primary respiratory chain NADH:quinone oxidoreductase of mycobacteria and nonenzymatic oxidation of reduced CFZ by O2 yielding CFZ and reactive oxygen species (ROS). This pathway was demonstrated using isolated membranes and purified recombinant NDH-2. The reduction and oxidation of CFZ was measured spectrally, and the production of ROS was measured using a coupled assay system with Amplex Red. Supporting the ROS-based killing mechanism, bacteria grown in the presence of antioxidants are more resistant to CFZ. CFZ-mediated increase in NADH oxidation and ROS production were not observed in membranes from three different Gram-negative bacteria but was observed in Staphylococcus aureus and Saccharomyces cerevisiae, which is consistent with the known antimicrobial specificity of CFZ. A more soluble analog of CFZ, KS6, was synthesized and was shown to have the same activities as CFZ. These studies describe a pathway for a continuous and high rate of reactive oxygen species production in Mycobacterium smegmatis treated with CFZ and a CFZ analog as well as evidence that cell death produced by these agents are related to the production of these radical species.</description><subject>Amyloid</subject><subject>Blood</subject><subject>Brain</subject><subject>RNA Binding Protein</subject><subject>RNA Folding</subject><subject>Translation Control</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><recordid>eNp1kM1LAzEQxYMoWKtnr_kHts0km-2ut1I_KmwtSgVvIR-zkNJuSrYV17_e1Hr1XYaBeY95P0JugY2ATfLx2tjRAtLGGZMMzsgAWCkyIeHjnAwY45BVXJaX5Krr1iwpr2BA6jd0B7v3oaWhobNNaPS33_oWqenporfBaLvH6PWGrvodUk5fpvfzu9eDb0M6Wn55F-JvhO7wmlw0etPhzd8ckvfHh9VsntXLp-fZtM4szwVksqhKDRwN5BWHRuqCS-PKAibOlDKJO4cGZWGgsFKUMBGapWaN5kI6ocWQjE-5Noaui9ioXfRbHXsFTB1hqARDHWGoE4zkqE4OTG99eoyqsx5bi85HtHvlgv_X-wMRkWS5</recordid><startdate>201103</startdate><enddate>201103</enddate><creator>Yano, Takahiro</creator><creator>Kassovska-Bratinova, Sacha</creator><creator>Teh, J. Shin</creator><creator>Winkler, Jeffrey</creator><creator>Sullivan, Kevin</creator><creator>Isaacs, Andre</creator><creator>Schechter, Norman M.</creator><creator>Rubin, Harvey</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>201103</creationdate><title>Reduction of Clofazimine by Mycobacterial Type 2 NADH:Quinone Oxidoreductase</title><author>Yano, Takahiro ; Kassovska-Bratinova, Sacha ; Teh, J. Shin ; Winkler, Jeffrey ; Sullivan, Kevin ; Isaacs, Andre ; Schechter, Norman M. ; Rubin, Harvey</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2431-5698a12eb14921f5a625bd8617db855552ddebe56b16c538173a0110fa235d3a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Amyloid</topic><topic>Blood</topic><topic>Brain</topic><topic>RNA Binding Protein</topic><topic>RNA Folding</topic><topic>Translation Control</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yano, Takahiro</creatorcontrib><creatorcontrib>Kassovska-Bratinova, Sacha</creatorcontrib><creatorcontrib>Teh, J. Shin</creatorcontrib><creatorcontrib>Winkler, Jeffrey</creatorcontrib><creatorcontrib>Sullivan, Kevin</creatorcontrib><creatorcontrib>Isaacs, Andre</creatorcontrib><creatorcontrib>Schechter, Norman M.</creatorcontrib><creatorcontrib>Rubin, Harvey</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>CrossRef</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yano, Takahiro</au><au>Kassovska-Bratinova, Sacha</au><au>Teh, J. Shin</au><au>Winkler, Jeffrey</au><au>Sullivan, Kevin</au><au>Isaacs, Andre</au><au>Schechter, Norman M.</au><au>Rubin, Harvey</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Reduction of Clofazimine by Mycobacterial Type 2 NADH:Quinone Oxidoreductase</atitle><jtitle>The Journal of biological chemistry</jtitle><date>2011-03</date><risdate>2011</risdate><volume>286</volume><issue>12</issue><spage>10276</spage><epage>10287</epage><pages>10276-10287</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The mechanism of action of clofazimine (CFZ), an antimycobacterial drug with a long history, is not well understood. The present study describes a redox cycling pathway that involves the enzymatic reduction of CFZ by NDH-2, the primary respiratory chain NADH:quinone oxidoreductase of mycobacteria and nonenzymatic oxidation of reduced CFZ by O2 yielding CFZ and reactive oxygen species (ROS). This pathway was demonstrated using isolated membranes and purified recombinant NDH-2. The reduction and oxidation of CFZ was measured spectrally, and the production of ROS was measured using a coupled assay system with Amplex Red. Supporting the ROS-based killing mechanism, bacteria grown in the presence of antioxidants are more resistant to CFZ. CFZ-mediated increase in NADH oxidation and ROS production were not observed in membranes from three different Gram-negative bacteria but was observed in Staphylococcus aureus and Saccharomyces cerevisiae, which is consistent with the known antimicrobial specificity of CFZ. A more soluble analog of CFZ, KS6, was synthesized and was shown to have the same activities as CFZ. These studies describe a pathway for a continuous and high rate of reactive oxygen species production in Mycobacterium smegmatis treated with CFZ and a CFZ analog as well as evidence that cell death produced by these agents are related to the production of these radical species.</abstract><pub>Elsevier Inc</pub><doi>10.1074/jbc.M110.200501</doi><oa>free_for_read</oa></addata></record>