X-ray Crystallography Reveals a Reduced Substrate Complex of UDP-Galactopyranose Mutase Poised for Covalent Catalysis by Flavin
The flavoenzyme uridine 5′-diphosphate galactopyranose mutase (UGM or Glf) catalyzes the interconversion of UDP-galactopyranose and UDP-galactofuranose. The latter is a key building block for cell wall construction in numerous pathogens, including Mycobacterium tuberculosis. Mechanistic studies of U...
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| Veröffentlicht in: | Biochemistry (Easton) 2009-10, Vol.48 (39), p.9171-9173 |
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| creator | Gruber, Todd D Westler, William M Kiessling, Laura L Forest, Katrina T |
| description | The flavoenzyme uridine 5′-diphosphate galactopyranose mutase (UGM or Glf) catalyzes the interconversion of UDP-galactopyranose and UDP-galactofuranose. The latter is a key building block for cell wall construction in numerous pathogens, including Mycobacterium tuberculosis. Mechanistic studies of UGM suggested a novel role for the flavin, and we previously provided evidence that the catalytic mechanism proceeds through a covalent flavin−galactose iminium. Here, we describe 2.3 and 2.5 Å resolution X-ray crystal structures of the substrate-bound enzyme in oxidized and reduced forms, respectively. In the latter, C1 of the substrate is 3.6 Å from the nucleophilic flavin N5 position. This orientation is consistent with covalent catalysis by flavin. |
| doi_str_mv | 10.1021/bi901437v |
| format | Article |
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(ANL), Argonne, IL (United States)</creatorcontrib><description>The flavoenzyme uridine 5′-diphosphate galactopyranose mutase (UGM or Glf) catalyzes the interconversion of UDP-galactopyranose and UDP-galactofuranose. The latter is a key building block for cell wall construction in numerous pathogens, including Mycobacterium tuberculosis. Mechanistic studies of UGM suggested a novel role for the flavin, and we previously provided evidence that the catalytic mechanism proceeds through a covalent flavin−galactose iminium. Here, we describe 2.3 and 2.5 Å resolution X-ray crystal structures of the substrate-bound enzyme in oxidized and reduced forms, respectively. In the latter, C1 of the substrate is 3.6 Å from the nucleophilic flavin N5 position. This orientation is consistent with covalent catalysis by flavin.</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi901437v</identifier><identifier>PMID: 19719175</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>CATALYSIS ; CELL WALL ; CONSTRUCTION ; CRYSTAL STRUCTURE ; CRYSTALLOGRAPHY ; Crystallography, X-Ray ; ENZYMES ; Flavin-Adenine Dinucleotide - chemistry ; Flavin-Adenine Dinucleotide - metabolism ; Intramolecular Transferases - chemistry ; Intramolecular Transferases - metabolism ; ISOALLOXAZINES ; Klebsiella pneumoniae - enzymology ; MATERIALS SCIENCE ; MYCOBACTERIUM TUBERCULOSIS ; ORIENTATION ; Oxidation-Reduction ; PATHOGENS ; RESOLUTION ; Substrate Specificity ; SUBSTRATES ; URIDINE</subject><ispartof>Biochemistry (Easton), 2009-10, Vol.48 (39), p.9171-9173</ispartof><rights>Copyright © 2009 American Chemical Society</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a463t-6bbc6ecaa88faa7855f7b89f798e2234a08a6877aee4b5a2164b6672e552e1483</citedby><cites>FETCH-LOGICAL-a463t-6bbc6ecaa88faa7855f7b89f798e2234a08a6877aee4b5a2164b6672e552e1483</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/bi901437v$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bi901437v$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>230,314,780,784,885,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19719175$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/1005941$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Gruber, Todd D</creatorcontrib><creatorcontrib>Westler, William M</creatorcontrib><creatorcontrib>Kiessling, Laura L</creatorcontrib><creatorcontrib>Forest, Katrina T</creatorcontrib><creatorcontrib>Argonne National Lab. (ANL), Argonne, IL (United States)</creatorcontrib><title>X-ray Crystallography Reveals a Reduced Substrate Complex of UDP-Galactopyranose Mutase Poised for Covalent Catalysis by Flavin</title><title>Biochemistry (Easton)</title><addtitle>Biochemistry</addtitle><description>The flavoenzyme uridine 5′-diphosphate galactopyranose mutase (UGM or Glf) catalyzes the interconversion of UDP-galactopyranose and UDP-galactofuranose. The latter is a key building block for cell wall construction in numerous pathogens, including Mycobacterium tuberculosis. Mechanistic studies of UGM suggested a novel role for the flavin, and we previously provided evidence that the catalytic mechanism proceeds through a covalent flavin−galactose iminium. Here, we describe 2.3 and 2.5 Å resolution X-ray crystal structures of the substrate-bound enzyme in oxidized and reduced forms, respectively. In the latter, C1 of the substrate is 3.6 Å from the nucleophilic flavin N5 position. This orientation is consistent with covalent catalysis by flavin.</description><subject>CATALYSIS</subject><subject>CELL WALL</subject><subject>CONSTRUCTION</subject><subject>CRYSTAL STRUCTURE</subject><subject>CRYSTALLOGRAPHY</subject><subject>Crystallography, X-Ray</subject><subject>ENZYMES</subject><subject>Flavin-Adenine Dinucleotide - chemistry</subject><subject>Flavin-Adenine Dinucleotide - metabolism</subject><subject>Intramolecular Transferases - chemistry</subject><subject>Intramolecular Transferases - metabolism</subject><subject>ISOALLOXAZINES</subject><subject>Klebsiella pneumoniae - enzymology</subject><subject>MATERIALS SCIENCE</subject><subject>MYCOBACTERIUM TUBERCULOSIS</subject><subject>ORIENTATION</subject><subject>Oxidation-Reduction</subject><subject>PATHOGENS</subject><subject>RESOLUTION</subject><subject>Substrate Specificity</subject><subject>SUBSTRATES</subject><subject>URIDINE</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkV9rFDEUxQdR7Lb64BeQIEjxYTSZyZ-Zl4KMtgoVi1rwLdzJ3ummZCdrkhmcJ7-6kV2qguDTTbi_e85NTlE8YfQloxV71duWMl6r-V6xYqKiJW9bcb9YUUplWbWSHhXHMd7mK6eKPyyOWKtYy5RYFT--lgEW0oUlJnDO3wTYbRbyCWcEFwnk03oyuCafpz6mAAlJ57c7h9-JH8j1m6vyAhyY5HdLgNFHJB-mBLlceRvz2OBDHpjB4ZhIB9ljiTaSfiHnDmY7PioeDNkIHx_qSXF9_vZL9668_Hjxvnt9WQKXdSpl3xuJBqBpBgDVCDGovmkH1TZYVTUH2oBslAJE3guomOS9lKpCISpkvKlPirO97m7qt7g2eZ0ATu-C3UJYtAer_-6MdqNv_KyrbJYdssCzvYCPyepobEKzMX4c0STNKBUtZxk6PbgE_23CmPTWRoPOwYh-iloJLjjlVfN_suZU5ox4Jl_sSRN8jAGHu6UZ1b_i13fxZ_bpn6_8TR7yzsDzPQAm6ls_hTF_-j-EfgLDy7hq</recordid><startdate>20091006</startdate><enddate>20091006</enddate><creator>Gruber, Todd D</creator><creator>Westler, William M</creator><creator>Kiessling, Laura L</creator><creator>Forest, Katrina T</creator><general>American Chemical Society</general><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><scope>7QL</scope><scope>C1K</scope><scope>OTOTI</scope><scope>5PM</scope></search><sort><creationdate>20091006</creationdate><title>X-ray Crystallography Reveals a Reduced Substrate Complex of UDP-Galactopyranose Mutase Poised for Covalent Catalysis by Flavin</title><author>Gruber, Todd D ; Westler, William M ; Kiessling, Laura L ; Forest, Katrina T</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a463t-6bbc6ecaa88faa7855f7b89f798e2234a08a6877aee4b5a2164b6672e552e1483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>CATALYSIS</topic><topic>CELL WALL</topic><topic>CONSTRUCTION</topic><topic>CRYSTAL STRUCTURE</topic><topic>CRYSTALLOGRAPHY</topic><topic>Crystallography, X-Ray</topic><topic>ENZYMES</topic><topic>Flavin-Adenine Dinucleotide - chemistry</topic><topic>Flavin-Adenine Dinucleotide - metabolism</topic><topic>Intramolecular Transferases - chemistry</topic><topic>Intramolecular Transferases - metabolism</topic><topic>ISOALLOXAZINES</topic><topic>Klebsiella pneumoniae - enzymology</topic><topic>MATERIALS SCIENCE</topic><topic>MYCOBACTERIUM TUBERCULOSIS</topic><topic>ORIENTATION</topic><topic>Oxidation-Reduction</topic><topic>PATHOGENS</topic><topic>RESOLUTION</topic><topic>Substrate Specificity</topic><topic>SUBSTRATES</topic><topic>URIDINE</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gruber, Todd D</creatorcontrib><creatorcontrib>Westler, William M</creatorcontrib><creatorcontrib>Kiessling, Laura L</creatorcontrib><creatorcontrib>Forest, Katrina T</creatorcontrib><creatorcontrib>Argonne National Lab. 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(ANL), Argonne, IL (United States)</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>X-ray Crystallography Reveals a Reduced Substrate Complex of UDP-Galactopyranose Mutase Poised for Covalent Catalysis by Flavin</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>2009-10-06</date><risdate>2009</risdate><volume>48</volume><issue>39</issue><spage>9171</spage><epage>9173</epage><pages>9171-9173</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>The flavoenzyme uridine 5′-diphosphate galactopyranose mutase (UGM or Glf) catalyzes the interconversion of UDP-galactopyranose and UDP-galactofuranose. The latter is a key building block for cell wall construction in numerous pathogens, including Mycobacterium tuberculosis. Mechanistic studies of UGM suggested a novel role for the flavin, and we previously provided evidence that the catalytic mechanism proceeds through a covalent flavin−galactose iminium. Here, we describe 2.3 and 2.5 Å resolution X-ray crystal structures of the substrate-bound enzyme in oxidized and reduced forms, respectively. In the latter, C1 of the substrate is 3.6 Å from the nucleophilic flavin N5 position. This orientation is consistent with covalent catalysis by flavin.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>19719175</pmid><doi>10.1021/bi901437v</doi><tpages>3</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | CATALYSIS CELL WALL CONSTRUCTION CRYSTAL STRUCTURE CRYSTALLOGRAPHY Crystallography, X-Ray ENZYMES Flavin-Adenine Dinucleotide - chemistry Flavin-Adenine Dinucleotide - metabolism Intramolecular Transferases - chemistry Intramolecular Transferases - metabolism ISOALLOXAZINES Klebsiella pneumoniae - enzymology MATERIALS SCIENCE MYCOBACTERIUM TUBERCULOSIS ORIENTATION Oxidation-Reduction PATHOGENS RESOLUTION Substrate Specificity SUBSTRATES URIDINE |
| title | X-ray Crystallography Reveals a Reduced Substrate Complex of UDP-Galactopyranose Mutase Poised for Covalent Catalysis by Flavin |
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