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[[Image:1r50.jpg|left|200px]]
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{{STRUCTURE_1r50|  PDB=1r50  |  SCENE=  }}
'''Bacillus subtilis lipase A with covalently bound Sc-IPG-phosphonate-inhibitor'''


==Bacillus subtilis lipase A with covalently bound Sc-IPG-phosphonate-inhibitor==
<StructureSection load='1r50' size='340' side='right'caption='[[1r50]], [[Resolution|resolution]] 1.45&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[1r50]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1R50 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1R50 FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.45&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=SIL:[(4S)-2,2-DIMETHYL-1,3-DIOXOLAN-4-YL]METHYL+HYDROGEN+HEX-5-ENYLPHOSPHONATE'>SIL</scene></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=1r50 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1r50 OCA], [https://pdbe.org/1r50 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1r50 RCSB], [https://www.ebi.ac.uk/pdbsum/1r50 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1r50 ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/ESTA_BACSU ESTA_BACSU] Active toward p-nitrophenyl esters and triacylglycerides with a marked preference for esters with C8 acyl groups.<ref>PMID:8396026</ref>
== Evolutionary Conservation ==
[[Image:Consurf_key_small.gif|200px|right]]
Check<jmol>
  <jmolCheckbox>
    <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/r5/1r50_consurf.spt"</scriptWhenChecked>
    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.spt</scriptWhenUnchecked>
    <text>to colour the structure by Evolutionary Conservation</text>
  </jmolCheckbox>
</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1r50 ConSurf].
<div style="clear:both"></div>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Phage display can be used as a protein-engineering tool for the selection of proteins with desirable binding properties from a library of mutants. Here we describe the application of this method for the directed evolution of Bacillus subtilis lipase A, an enzyme that has important properties for the preparation of the pharmaceutically relevant chiral compound 1,2-O-isopropylidene-sn-glycerol (IPG). PCR mutagenesis with spiked oligonucleotides was employed for saturation mutagenesis of a stretch of amino acids near the active site. After expression of these mutants on bacteriophages, dual selection with (S)-(+)- and (R)-(-)-IPG stereoisomers covalently coupled to enantiomeric phosphonate suicide inhibitors (SIRAN Sc and Rc inhibitors, respectively) was used for the isolation of variants with inverted enantioselectivity. The mutants were further characterised by determination of their Michaelis-Menten parameters. The 3D structures of the Sc and Rc inhibitor-lipase complexes were determined and provided structural insight into the mechanism of enantioselectivity of the enzyme. In conclusion, we have used phage display as a fast and reproducible method for the selection of Bacillus lipase A mutant enzymes with inverted enantioselectivity.


==Overview==
Directed evolution of Bacillus subtilis lipase A by use of enantiomeric phosphonate inhibitors: crystal structures and phage display selection.,Droge MJ, Boersma YL, van Pouderoyen G, Vrenken TE, Ruggeberg CJ, Reetz MT, Dijkstra BW, Quax WJ Chembiochem. 2006 Jan;7(1):149-57. PMID:16342303<ref>PMID:16342303</ref>
Phage display can be used as a protein-engineering tool for the selection of proteins with desirable binding properties from a library of mutants. Here we describe the application of this method for the directed evolution of Bacillus subtilis lipase A, an enzyme that has important properties for the preparation of the pharmaceutically relevant chiral compound 1,2-O-isopropylidene-sn-glycerol (IPG). PCR mutagenesis with spiked oligonucleotides was employed for saturation mutagenesis of a stretch of amino acids near the active site. After expression of these mutants on bacteriophages, dual selection with (S)-(+)- and (R)-(-)-IPG stereoisomers covalently coupled to enantiomeric phosphonate suicide inhibitors (SIRAN Sc and Rc inhibitors, respectively) was used for the isolation of variants with inverted enantioselectivity. The mutants were further characterised by determination of their Michaelis-Menten parameters. The 3D structures of the Sc and Rc inhibitor-lipase complexes were determined and provided structural insight into the mechanism of enantioselectivity of the enzyme. In conclusion, we have used phage display as a fast and reproducible method for the selection of Bacillus lipase A mutant enzymes with inverted enantioselectivity.


==About this Structure==
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
1R50 is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1R50 OCA].
</div>
<div class="pdbe-citations 1r50" style="background-color:#fffaf0;"></div>


==Reference==
==See Also==
Directed evolution of Bacillus subtilis lipase A by use of enantiomeric phosphonate inhibitors: crystal structures and phage display selection., Droge MJ, Boersma YL, van Pouderoyen G, Vrenken TE, Ruggeberg CJ, Reetz MT, Dijkstra BW, Quax WJ, Chembiochem. 2006 Jan;7(1):149-57. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16342303 16342303]
*[[Lipase 3D Structures|Lipase 3D Structures]]
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Bacillus subtilis]]
[[Category: Bacillus subtilis]]
[[Category: Single protein]]
[[Category: Large Structures]]
[[Category: Triacylglycerol lipase]]
[[Category: Dijkstra BW]]
[[Category: Dijkstra, B W.]]
[[Category: Droege MJ]]
[[Category: Droege, M J.]]
[[Category: Quax WJ]]
[[Category: Pouderoyen, G Van.]]
[[Category: Reetz MT]]
[[Category: Quax, W J.]]
[[Category: Rueggeberg CJ]]
[[Category: Reetz, M T.]]
[[Category: Van Pouderoyen G]]
[[Category: Rueggeberg, C J.]]
[[Category: Vrenken TE]]
[[Category: Vrenken, T E.]]
[[Category: Alpha/beta hydrolase]]
[[Category: Lipase]]
[[Category: Phosphonate inhibitor]]
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May  3 07:05:00 2008''

Latest revision as of 07:25, 9 October 2024

Bacillus subtilis lipase A with covalently bound Sc-IPG-phosphonate-inhibitor

1r50, resolution 1.45Å

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