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| | ==Crystal Structure of B. thuringiensis AiiA mutant F107W with N-decanoyl-L-homoserine bound at the active site== | | ==Crystal Structure of B. thuringiensis AiiA mutant F107W with N-decanoyl-L-homoserine bound at the active site== |
| - | <StructureSection load='4j5h' size='340' side='right' caption='[[4j5h]], [[Resolution|resolution]] 1.45Å' scene=''> | + | <StructureSection load='4j5h' size='340' side='right'caption='[[4j5h]], [[Resolution|resolution]] 1.45Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4j5h]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/"bacillus_cereus_var._thuringiensis"_smith_et_al._1952 "bacillus cereus var. thuringiensis" smith et al. 1952]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4J5H OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4J5H FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4j5h]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Bacillus_thuringiensis Bacillus thuringiensis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4J5H OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4J5H FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=1K4:N-DECANOYL-L-HOMOSERINE'>1K4</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr> | + | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=1K4:N-DECANOYL-L-HOMOSERINE'>1K4</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4j5f|4j5f]]</td></tr>
| + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=4j5h FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4j5h OCA], [https://pdbe.org/4j5h PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4j5h RCSB], [https://www.ebi.ac.uk/pdbsum/4j5h PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4j5h ProSAT]</span></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">aiiA ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1428 "Bacillus cereus var. thuringiensis" Smith et al. 1952])</td></tr>
| + | |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Quorum-quenching_N-acyl-homoserine_lactonase Quorum-quenching N-acyl-homoserine lactonase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.1.81 3.1.1.81] </span></td></tr>
| + | |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4j5h FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4j5h OCA], [http://pdbe.org/4j5h PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4j5h RCSB], [http://www.ebi.ac.uk/pdbsum/4j5h PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4j5h ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/AHLL_BACTU AHLL_BACTU]] Catalyzes hydrolysis of N-hexanoyl-(S)-homoserine lactone, but not the R-enantiomer. Hydrolyzes short- and long-chain N-acyl homoserine lactones with or without 3-oxo substitution at C3, has maximum activity on C10-AHL.<ref>PMID:15895999</ref> | + | [https://www.uniprot.org/uniprot/AHLL_BACTU AHLL_BACTU] Catalyzes hydrolysis of N-hexanoyl-(S)-homoserine lactone, but not the R-enantiomer. Hydrolyzes short- and long-chain N-acyl homoserine lactones with or without 3-oxo substitution at C3, has maximum activity on C10-AHL.<ref>PMID:15895999</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Bacillus cereus var. thuringiensis smith et al. 1952]] | + | [[Category: Bacillus thuringiensis]] |
| - | [[Category: Quorum-quenching N-acyl-homoserine lactonase]] | + | [[Category: Large Structures]] |
| - | [[Category: Fast, W]] | + | [[Category: Fast W]] |
| - | [[Category: Lajoie, A]] | + | [[Category: Lajoie A]] |
| - | [[Category: Liu, C F]] | + | [[Category: Liu CF]] |
| - | [[Category: Liu, D]] | + | [[Category: Liu D]] |
| - | [[Category: Momb, J]] | + | [[Category: Momb J]] |
| - | [[Category: Petsko, G A]] | + | [[Category: Petsko GA]] |
| - | [[Category: Ringe, D]] | + | [[Category: Ringe D]] |
| - | [[Category: Thomas, P W]] | + | [[Category: Thomas PW]] |
| - | [[Category: Aiia]]
| + | |
| - | [[Category: Beta-hairpin loop]]
| + | |
| - | [[Category: Dizinc hydrolase]]
| + | |
| - | [[Category: Hydrolase-hydrolase substrate complex]]
| + | |
| - | [[Category: Lactonase]]
| + | |
| - | [[Category: N-acyl homoserine lactone]]
| + | |
| - | [[Category: Quorum quenching]]
| + | |
| - | [[Category: Substrate specificity]]
| + | |
| Structural highlights
Function
AHLL_BACTU Catalyzes hydrolysis of N-hexanoyl-(S)-homoserine lactone, but not the R-enantiomer. Hydrolyzes short- and long-chain N-acyl homoserine lactones with or without 3-oxo substitution at C3, has maximum activity on C10-AHL.[1]
Publication Abstract from PubMed
Autoinducer inactivator A (AiiA) is a metal-dependent N-acyl homoserine lactone hydrolase that displays broad substrate specificity but shows a preference for substrates with long N-acyl substitutions. Previously, crystal structures of AiiA in complex with the ring-opened product N-hexanoyl-l-homoserine revealed binding interactions near the metal center but did not identify a binding pocket for the N-acyl chains of longer substrates. Here we report the crystal structure of an AiiA mutant, F107W, determined in the presence and absence of N-decanoyl-l-homoserine. F107 is located in a hydrophobic cavity adjacent to the previously identified ligand binding pocket, and the F107W mutation results in the formation of an unexpected interaction with the ring-opened product. Notably, the structure reveals a previously unidentified hydrophobic binding pocket for the substrate's N-acyl chain. Two aromatic residues, F64 and F68, form a hydrophobic clamp, centered around the seventh carbon in the product-bound structure's decanoyl chain, making an interaction that would also be available for longer substrates, but not for shorter substrates. Steady-state kinetics using substrates of various lengths with AiiA bearing mutations at the hydrophobic clamp, including insertion of a redox-sensitive cysteine pair, confirms the importance of this hydrophobic feature for substrate preference. Identifying the specificity determinants of AiiA will aid the development of more selective quorum-quenching enzymes as tools and as potential therapeutics.
A phenylalanine clamp controls substrate specificity in the quorum-quenching metallo-gamma-lactonase from Bacillus thuringiensis.,Liu CF, Liu D, Momb J, Thomas PW, Lajoie A, Petsko GA, Fast W, Ringe D Biochemistry. 2013 Mar 5;52(9):1603-10. doi: 10.1021/bi400050j. Epub 2013 Feb 20. PMID:23387521[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Thomas PW, Stone EM, Costello AL, Tierney DL, Fast W. The quorum-quenching lactonase from Bacillus thuringiensis is a metalloprotein. Biochemistry. 2005 May 24;44(20):7559-69. PMID:15895999 doi:http://dx.doi.org/10.1021/bi050050m
- ↑ Liu CF, Liu D, Momb J, Thomas PW, Lajoie A, Petsko GA, Fast W, Ringe D. A phenylalanine clamp controls substrate specificity in the quorum-quenching metallo-gamma-lactonase from Bacillus thuringiensis. Biochemistry. 2013 Mar 5;52(9):1603-10. doi: 10.1021/bi400050j. Epub 2013 Feb 20. PMID:23387521 doi:10.1021/bi400050j
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