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Crystal structure of UDP- N-acetylglucosamine enolpyruvyltransferase, the target of the antibiotic fosfomycin

Background The ever increasing number of antibiotic resistant bacteria has fuelled interest in the development of new antibiotics and other antibacterial agents. The major structural element of the bacterial cell wall is the heteropolymer peptidoglycan and the enzymes of peptidoglycan biosynthesis a...

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Published in:Structure (London) 1996-09, Vol.4 (9), p.1065-1075
Main Authors: Schönbrunn, Ernst, Sack, Stefan, Eschenburg, Susanne, Perrakis, Anastassis, Krekel, Florian, Amrhein, Nikolaus, Mandelkow, Eckhard
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cited_by cdi_FETCH-LOGICAL-c502t-fec972c4f54b905e6745c0a6b7c2d873fd7670e968c0b0a162d8a86cf22a16313
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container_issue 9
container_start_page 1065
container_title Structure (London)
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creator Schönbrunn, Ernst
Sack, Stefan
Eschenburg, Susanne
Perrakis, Anastassis
Krekel, Florian
Amrhein, Nikolaus
Mandelkow, Eckhard
description Background The ever increasing number of antibiotic resistant bacteria has fuelled interest in the development of new antibiotics and other antibacterial agents. The major structural element of the bacterial cell wall is the heteropolymer peptidoglycan and the enzymes of peptidoglycan biosynthesis are potential targets for antibacterial agents. One such enzyme is UDP- N-acetylglucosamine enolpyruvyltransferase (EPT) which catalyzes the first committed step in peptidoglycan biosynthesis: the transfer of the enolpyruvyl moiety of phosphoenolpyruvate (PEP) to the 3-hydroxyl of UDP- N-acetylglucosamine (UDPGlcNAc). EPT is of potential pharmaceutical interest because it is inhibited by the broad spectrum antibiotic fosfomycin. Results The crystal structure of substrate-free EPT has been determined at 2.0 å resolution. The structure reveals a two-domain protein with an unusual fold (inside out α/ β barrel) which is built up from the sixfold repetition of one folding unit. The only repetitive element in the amino acid sequence is a short motif, Leu-X 3-Gly(Ala), which is responsible for the formation of hydrogen-bond interactions between the folding units. An enzyme which catalyzes a similar reaction to EPT, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), has a very similar structure despite an amino acid sequence identity of only 25%. To date, only these two enzymes appear to display this characteristic fold. Conclusions The present structure reflects the open conformation of the enzyme which is probably stabilized through two residues, a lysine and an arginine, located in the cleft between the domains. Binding of the negatively charged UDPGlcNAc to these residues could neutralize the repulsive force between the two domains, thereby allowing the movement of a catalytically active cysteine residue towards the cleft.
doi_str_mv 10.1016/S0969-2126(96)00113-X
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The major structural element of the bacterial cell wall is the heteropolymer peptidoglycan and the enzymes of peptidoglycan biosynthesis are potential targets for antibacterial agents. One such enzyme is UDP- N-acetylglucosamine enolpyruvyltransferase (EPT) which catalyzes the first committed step in peptidoglycan biosynthesis: the transfer of the enolpyruvyl moiety of phosphoenolpyruvate (PEP) to the 3-hydroxyl of UDP- N-acetylglucosamine (UDPGlcNAc). EPT is of potential pharmaceutical interest because it is inhibited by the broad spectrum antibiotic fosfomycin. Results The crystal structure of substrate-free EPT has been determined at 2.0 å resolution. The structure reveals a two-domain protein with an unusual fold (inside out α/ β barrel) which is built up from the sixfold repetition of one folding unit. The only repetitive element in the amino acid sequence is a short motif, Leu-X 3-Gly(Ala), which is responsible for the formation of hydrogen-bond interactions between the folding units. An enzyme which catalyzes a similar reaction to EPT, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), has a very similar structure despite an amino acid sequence identity of only 25%. To date, only these two enzymes appear to display this characteristic fold. Conclusions The present structure reflects the open conformation of the enzyme which is probably stabilized through two residues, a lysine and an arginine, located in the cleft between the domains. Binding of the negatively charged UDPGlcNAc to these residues could neutralize the repulsive force between the two domains, thereby allowing the movement of a catalytically active cysteine residue towards the cleft.</description><identifier>ISSN: 0969-2126</identifier><identifier>EISSN: 1878-4186</identifier><identifier>DOI: 10.1016/S0969-2126(96)00113-X</identifier><identifier>PMID: 8805592</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Alkyl and Aryl Transferases ; Amino Acid Sequence ; Catalysis ; Crystallography, X-Ray ; domain movement ; folding ; Fosfomycin - metabolism ; hinge ; Molecular Sequence Data ; peptidoglycan biosynthesis ; Protein Conformation ; Sequence Homology, Amino Acid ; sequence motif ; Substrate Specificity ; Transferases - chemistry ; Transferases - metabolism</subject><ispartof>Structure (London), 1996-09, Vol.4 (9), p.1065-1075</ispartof><rights>1996 Elsevier Science Ltd</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c502t-fec972c4f54b905e6745c0a6b7c2d873fd7670e968c0b0a162d8a86cf22a16313</citedby><cites>FETCH-LOGICAL-c502t-fec972c4f54b905e6745c0a6b7c2d873fd7670e968c0b0a162d8a86cf22a16313</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8805592$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Schönbrunn, Ernst</creatorcontrib><creatorcontrib>Sack, Stefan</creatorcontrib><creatorcontrib>Eschenburg, Susanne</creatorcontrib><creatorcontrib>Perrakis, Anastassis</creatorcontrib><creatorcontrib>Krekel, Florian</creatorcontrib><creatorcontrib>Amrhein, Nikolaus</creatorcontrib><creatorcontrib>Mandelkow, Eckhard</creatorcontrib><title>Crystal structure of UDP- N-acetylglucosamine enolpyruvyltransferase, the target of the antibiotic fosfomycin</title><title>Structure (London)</title><addtitle>Structure</addtitle><description>Background The ever increasing number of antibiotic resistant bacteria has fuelled interest in the development of new antibiotics and other antibacterial agents. 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The only repetitive element in the amino acid sequence is a short motif, Leu-X 3-Gly(Ala), which is responsible for the formation of hydrogen-bond interactions between the folding units. An enzyme which catalyzes a similar reaction to EPT, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), has a very similar structure despite an amino acid sequence identity of only 25%. To date, only these two enzymes appear to display this characteristic fold. Conclusions The present structure reflects the open conformation of the enzyme which is probably stabilized through two residues, a lysine and an arginine, located in the cleft between the domains. Binding of the negatively charged UDPGlcNAc to these residues could neutralize the repulsive force between the two domains, thereby allowing the movement of a catalytically active cysteine residue towards the cleft.</description><subject>Alkyl and Aryl Transferases</subject><subject>Amino Acid Sequence</subject><subject>Catalysis</subject><subject>Crystallography, X-Ray</subject><subject>domain movement</subject><subject>folding</subject><subject>Fosfomycin - metabolism</subject><subject>hinge</subject><subject>Molecular Sequence Data</subject><subject>peptidoglycan biosynthesis</subject><subject>Protein Conformation</subject><subject>Sequence Homology, Amino Acid</subject><subject>sequence motif</subject><subject>Substrate Specificity</subject><subject>Transferases - chemistry</subject><subject>Transferases - metabolism</subject><issn>0969-2126</issn><issn>1878-4186</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><recordid>eNqFkE9r3DAQxUVJSTdpP0LAp5BCnIxkW5ZPoWz_JBDaQhvITcjjUapiWxtJDvjbx5tdcs1pmJn35jE_xk44XHDg8vIPNLLJBRfyrJGfATgv8vt3bMVVrfKSK3nAVq-SD-woxv8AICqAQ3aoFFRVI1ZsWIc5JtNnMYUJ0xQo8za7-_o7z37mBinN_UM_oY9mcCNlNPp-M4fpae5TMGO0FEyk8yz9oyyZ8EBpa992ZkyudT45zKyP1g8zuvEje29NH-nTvh6zu-_f_q6v89tfP27WX25zrECk3BI2tcDSVmXbQEWyLisEI9saRafqwna1rIEaqRBaMFwuU6MkWiGWpuDFMTvd3d0E_zhRTHpwEanvzUh-irpWBS9LKRZhtRNi8DEGsnoT3GDCrDnoLWb9gllvGepG6hfM-n7xnewDpnag7tW157rsr3Z7Wr58chR0REcjUucCYdKdd28kPAN7Io8C</recordid><startdate>19960915</startdate><enddate>19960915</enddate><creator>Schönbrunn, Ernst</creator><creator>Sack, Stefan</creator><creator>Eschenburg, Susanne</creator><creator>Perrakis, Anastassis</creator><creator>Krekel, Florian</creator><creator>Amrhein, Nikolaus</creator><creator>Mandelkow, Eckhard</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</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>19960915</creationdate><title>Crystal structure of UDP- N-acetylglucosamine enolpyruvyltransferase, the target of the antibiotic fosfomycin</title><author>Schönbrunn, Ernst ; Sack, Stefan ; Eschenburg, Susanne ; Perrakis, Anastassis ; Krekel, Florian ; Amrhein, Nikolaus ; Mandelkow, Eckhard</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c502t-fec972c4f54b905e6745c0a6b7c2d873fd7670e968c0b0a162d8a86cf22a16313</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Alkyl and Aryl Transferases</topic><topic>Amino Acid Sequence</topic><topic>Catalysis</topic><topic>Crystallography, X-Ray</topic><topic>domain movement</topic><topic>folding</topic><topic>Fosfomycin - metabolism</topic><topic>hinge</topic><topic>Molecular Sequence Data</topic><topic>peptidoglycan biosynthesis</topic><topic>Protein Conformation</topic><topic>Sequence Homology, Amino Acid</topic><topic>sequence motif</topic><topic>Substrate Specificity</topic><topic>Transferases - chemistry</topic><topic>Transferases - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Schönbrunn, Ernst</creatorcontrib><creatorcontrib>Sack, Stefan</creatorcontrib><creatorcontrib>Eschenburg, Susanne</creatorcontrib><creatorcontrib>Perrakis, Anastassis</creatorcontrib><creatorcontrib>Krekel, Florian</creatorcontrib><creatorcontrib>Amrhein, Nikolaus</creatorcontrib><creatorcontrib>Mandelkow, Eckhard</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</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>Structure (London)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Schönbrunn, Ernst</au><au>Sack, Stefan</au><au>Eschenburg, Susanne</au><au>Perrakis, Anastassis</au><au>Krekel, Florian</au><au>Amrhein, Nikolaus</au><au>Mandelkow, Eckhard</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Crystal structure of UDP- N-acetylglucosamine enolpyruvyltransferase, the target of the antibiotic fosfomycin</atitle><jtitle>Structure (London)</jtitle><addtitle>Structure</addtitle><date>1996-09-15</date><risdate>1996</risdate><volume>4</volume><issue>9</issue><spage>1065</spage><epage>1075</epage><pages>1065-1075</pages><issn>0969-2126</issn><eissn>1878-4186</eissn><abstract>Background The ever increasing number of antibiotic resistant bacteria has fuelled interest in the development of new antibiotics and other antibacterial agents. 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The only repetitive element in the amino acid sequence is a short motif, Leu-X 3-Gly(Ala), which is responsible for the formation of hydrogen-bond interactions between the folding units. An enzyme which catalyzes a similar reaction to EPT, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), has a very similar structure despite an amino acid sequence identity of only 25%. To date, only these two enzymes appear to display this characteristic fold. Conclusions The present structure reflects the open conformation of the enzyme which is probably stabilized through two residues, a lysine and an arginine, located in the cleft between the domains. 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identifier ISSN: 0969-2126
ispartof Structure (London), 1996-09, Vol.4 (9), p.1065-1075
issn 0969-2126
1878-4186
language eng
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source BACON - Elsevier - GLOBAL_SCIENCEDIRECT-OPENACCESS
subjects Alkyl and Aryl Transferases
Amino Acid Sequence
Catalysis
Crystallography, X-Ray
domain movement
folding
Fosfomycin - metabolism
hinge
Molecular Sequence Data
peptidoglycan biosynthesis
Protein Conformation
Sequence Homology, Amino Acid
sequence motif
Substrate Specificity
Transferases - chemistry
Transferases - metabolism
title Crystal structure of UDP- N-acetylglucosamine enolpyruvyltransferase, the target of the antibiotic fosfomycin
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