Loading…
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...
Saved in:
Published in: | Structure (London) 1996-09, Vol.4 (9), p.1065-1075 |
---|---|
Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
cited_by | cdi_FETCH-LOGICAL-c502t-fec972c4f54b905e6745c0a6b7c2d873fd7670e968c0b0a162d8a86cf22a16313 |
---|---|
cites | cdi_FETCH-LOGICAL-c502t-fec972c4f54b905e6745c0a6b7c2d873fd7670e968c0b0a162d8a86cf22a16313 |
container_end_page | 1075 |
container_issue | 9 |
container_start_page | 1065 |
container_title | Structure (London) |
container_volume | 4 |
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 |
format | article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_78314462</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S096921269600113X</els_id><sourcerecordid>78314462</sourcerecordid><originalsourceid>FETCH-LOGICAL-c502t-fec972c4f54b905e6745c0a6b7c2d873fd7670e968c0b0a162d8a86cf22a16313</originalsourceid><addsrcrecordid>eNqFkE9r3DAQxUVJSTdpP0LAp5BCnIxkW5ZPoWz_JBDaQhvITcjjUapiWxtJDvjbx5tdcs1pmJn35jE_xk44XHDg8vIPNLLJBRfyrJGfATgv8vt3bMVVrfKSK3nAVq-SD-woxv8AICqAQ3aoFFRVI1ZsWIc5JtNnMYUJ0xQo8za7-_o7z37mBinN_UM_oY9mcCNlNPp-M4fpae5TMGO0FEyk8yz9oyyZ8EBpa992ZkyudT45zKyP1g8zuvEje29NH-nTvh6zu-_f_q6v89tfP27WX25zrECk3BI2tcDSVmXbQEWyLisEI9saRafqwna1rIEaqRBaMFwuU6MkWiGWpuDFMTvd3d0E_zhRTHpwEanvzUh-irpWBS9LKRZhtRNi8DEGsnoT3GDCrDnoLWb9gllvGepG6hfM-n7xnewDpnag7tW157rsr3Z7Wr58chR0REcjUucCYdKdd28kPAN7Io8C</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>78314462</pqid></control><display><type>article</type><title>Crystal structure of UDP- N-acetylglucosamine enolpyruvyltransferase, the target of the antibiotic fosfomycin</title><source>BACON - Elsevier - GLOBAL_SCIENCEDIRECT-OPENACCESS</source><creator>Schönbrunn, Ernst ; Sack, Stefan ; Eschenburg, Susanne ; Perrakis, Anastassis ; Krekel, Florian ; Amrhein, Nikolaus ; Mandelkow, Eckhard</creator><creatorcontrib>Schönbrunn, Ernst ; Sack, Stefan ; Eschenburg, Susanne ; Perrakis, Anastassis ; Krekel, Florian ; Amrhein, Nikolaus ; Mandelkow, Eckhard</creatorcontrib><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.</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. 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><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. 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.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>8805592</pmid><doi>10.1016/S0969-2126(96)00113-X</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0969-2126 |
ispartof | Structure (London), 1996-09, Vol.4 (9), p.1065-1075 |
issn | 0969-2126 1878-4186 |
language | eng |
recordid | cdi_proquest_miscellaneous_78314462 |
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 |
url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-22T22%3A31%3A28IST&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=Crystal%20structure%20of%20UDP-%20N-acetylglucosamine%20enolpyruvyltransferase,%20the%20target%20of%20the%20antibiotic%20fosfomycin&rft.jtitle=Structure%20(London)&rft.au=Sch%C3%B6nbrunn,%20Ernst&rft.date=1996-09-15&rft.volume=4&rft.issue=9&rft.spage=1065&rft.epage=1075&rft.pages=1065-1075&rft.issn=0969-2126&rft.eissn=1878-4186&rft_id=info:doi/10.1016/S0969-2126(96)00113-X&rft_dat=%3Cproquest_cross%3E78314462%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c502t-fec972c4f54b905e6745c0a6b7c2d873fd7670e968c0b0a162d8a86cf22a16313%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=78314462&rft_id=info:pmid/8805592&rfr_iscdi=true |