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4av7

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Structure determination of the double mutant S233Y F250G from the sec- alkyl sulfatase PisA1

Structural highlights

4av7 is a 6 chain structure with sequence from Pseudomonas sp. DSM 6611. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 3Å
Ligands:	,
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

RSAKS_PSESP Alkylsulfatase that catalyzes the enantioselective hydrolysis of secondary-alkylsulfates with strict inversion of configuration, leading to the formation of homochiral (S)-configurated alcohols and nonreacted sulfate esters (PubMed:21770430, Ref.2, PubMed:23061549). The substrate spectrum includes a range of linear, branched or cyclic sec-alkylsulfates (PubMed:21770430). Can use sec-alkylsulfate esters bearing aromatic, olefinic and acetylenic moieties (Ref.2). Acts by cleaving the C-O bond, resulting in inversion at the carbon (PubMed:23061549).^[1] ^[2] ^[3]

Publication Abstract from PubMed

A highly enantioselective and stereoselective secondary alkylsulfatase from Pseudomonas sp. DSM6611 (Pisa1) was heterologously expressed in Escherichia coli BL21, and purified to homogeneity for kinetic and structural studies. Structure determination of Pisa1 by X-ray crystallography showed that the protein belongs to the family of metallo-beta-lactamases with a conserved binuclear Zn(2+) cluster in the active site. In contrast to a closely related alkylsulfatase from Pseudomonas aeruginosa (SdsA1), Pisa1 showed a preference for secondary rather than primary alkyl sulfates, and enantioselectively hydrolyzed the (R)-enantiomer of rac-2-octyl sulfate, yielding (S)-2-octanol with inversion of absolute configuration as a result of C-O bond cleavage. In order to elucidate the mechanism of inverting sulfate ester hydrolysis, for which no counterpart in chemical catalysis exists, we designed variants of Pisa1 guided by three-dimensional structure and docking experiments. In the course of these studies, we identified an invariant histidine (His317) near the sulfate-binding site as the general acid for crucial protonation of the sulfate leaving group. Additionally, amino acid replacements in the alkyl chain-binding pocket generated an enzyme variant that lost its stereoselectivity towards rac-2-octyl sulfate. These findings are discussed in light of the potential use of this enzyme family for applications in biocatalysis. DATABASE: The atomic coordinates and structural factors have been deposited in the Protein Data Bank under the accession codes 2YHE (wild type, crystal form I), 4AV7 (double variant Ser233-->Tyr/Ph250-->Gly) and 4AXH (wild type, crystal form II) STRUCTURED DIGITAL ABSTRACT: Pisa1 and Pisa1 bind by x-ray crystallography (View interaction).

Structure and mechanism of an inverting alkylsulfatase from Pseudomonas sp. DSM6611 specific for secondary alkyl sulfates.,Knaus T, Schober M, Kepplinger B, Faccinelli M, Pitzer J, Faber K, Macheroux P, Wagner U FEBS J. 2012 Dec;279(23):4374-84. doi: 10.1111/febs.12027. Epub 2012 Nov 7. PMID:23061549^[4]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Schober M, Gadler P, Knaus T, Kayer H, Birner-Grünberger R, Gülly C, Macheroux P, Wagner U, Faber K. A stereoselective inverting sec-alkylsulfatase for the deracemization of sec-alcohols. Org Lett. 2011 Aug 19;13(16):4296-9. PMID:21770430 doi:10.1021/ol201635y
↑ Knaus T, Schober M, Kepplinger B, Faccinelli M, Pitzer J, Faber K, Macheroux P, Wagner U. Structure and mechanism of an inverting alkylsulfatase from Pseudomonas sp. DSM6611 specific for secondary alkyl sulfates. FEBS J. 2012 Dec;279(23):4374-84. doi: 10.1111/febs.12027. Epub 2012 Nov 7. PMID:23061549 doi:http://dx.doi.org/10.1111/febs.12027
↑ Schober M, Gadler P, Knaus T, Kayer H, Birner-Grünberger R, Gülly C, Macheroux P, Wagner U, Faber K. A stereoselective inverting sec-alkylsulfatase for the deracemization of sec-alcohols. Org Lett. 2011 Aug 19;13(16):4296-9. PMID:21770430 doi:10.1021/ol201635y
↑ Knaus T, Schober M, Kepplinger B, Faccinelli M, Pitzer J, Faber K, Macheroux P, Wagner U. Structure and mechanism of an inverting alkylsulfatase from Pseudomonas sp. DSM6611 specific for secondary alkyl sulfates. FEBS J. 2012 Dec;279(23):4374-84. doi: 10.1111/febs.12027. Epub 2012 Nov 7. PMID:23061549 doi:http://dx.doi.org/10.1111/febs.12027

1 Structural highlights
2 Function
3 Publication Abstract from PubMed
4 See Also
5 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/4av7"

@@ Line 3: / Line 3: @@
 <StructureSection load='4av7' size='340' side='right'caption='[[4av7]], [[Resolution|resolution]] 3.00&Aring;' scene=''>
 == Structural highlights ==
-<table><tr><td colspan='2'>[[4av7]] is a 6 chain structure with sequence from [http://en.wikipedia.org/wiki/Pseudomonas_sp._dsm_6611 Pseudomonas sp. dsm 6611]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4AV7 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4AV7 FirstGlance]. <br>
+<table><tr><td colspan='2'>[[4av7]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_sp._DSM_6611 Pseudomonas sp. DSM 6611]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4AV7 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4AV7 FirstGlance]. <br>
-</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
+</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 3&#8491;</td></tr>
-<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[2yhe|2yhe]], [[4axh|4axh]]</td></tr>
+<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></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=4av7 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4av7 OCA], [http://pdbe.org/4av7 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4av7 RCSB], [http://www.ebi.ac.uk/pdbsum/4av7 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4av7 ProSAT]</span></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=4av7 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4av7 OCA], [https://pdbe.org/4av7 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4av7 RCSB], [https://www.ebi.ac.uk/pdbsum/4av7 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4av7 ProSAT]</span></td></tr>
 </table>
+== Function ==
+[https://www.uniprot.org/uniprot/RSAKS_PSESP RSAKS_PSESP] Alkylsulfatase that catalyzes the enantioselective hydrolysis of secondary-alkylsulfates with strict inversion of configuration, leading to the formation of homochiral (S)-configurated alcohols and nonreacted sulfate esters (PubMed:21770430, Ref.2, PubMed:23061549). The substrate spectrum includes a range of linear, branched or cyclic sec-alkylsulfates (PubMed:21770430). Can use sec-alkylsulfate esters bearing aromatic, olefinic and acetylenic moieties (Ref.2). Acts by cleaving the C-O bond, resulting in inversion at the carbon (PubMed:23061549).<ref>PMID:21770430</ref> <ref>PMID:23061549</ref> <ref>PMID:21770430</ref>
 <div style="background-color:#fffaf0;">
 == Publication Abstract from PubMed ==
@@ Line 17: / Line 19: @@
 </div>
 <div class="pdbe-citations 4av7" style="background-color:#fffaf0;"></div>
+==See Also==
+*[[Sulfatase 3D structures|Sulfatase 3D structures]]
 == References ==
 <references/>
 __TOC__
 </StructureSection>
-[[Category: Pseudomonas sp. dsm 6611]]
+[[Category: Large Structures]]
-[[Category: Faber, K]]
+[[Category: Pseudomonas sp. DSM 6611]]
-[[Category: Knaus, T]]
+[[Category: Faber K]]
-[[Category: Macharaux, P]]
+[[Category: Knaus T]]
-[[Category: Schober, M]]
+[[Category: Macharaux P]]
-[[Category: Wagner, U]]
+[[Category: Schober M]]
-[[Category: Double mutant]]
+[[Category: Wagner U]]
-[[Category: Hydrolase]]
-[[Category: Inverting alkylsulfatase]]

4av7

From Proteopedia

Current revision

Structure determination of the double mutant S233Y F250G from the sec- alkyl sulfatase PisA1

Structural highlights

Function

Publication Abstract from PubMed

See Also

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

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