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==Crystal structure of cold-active esterase PMGL3 from permafrost metagenomic library== | ==Crystal structure of cold-active esterase PMGL3 from permafrost metagenomic library== | ||
<StructureSection load='7b1x' size='340' side='right'caption='[[7b1x]]' scene=''> | <StructureSection load='7b1x' size='340' side='right'caption='[[7b1x]], [[Resolution|resolution]] 2.30Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7B1X OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7B1X FirstGlance]. <br> | <table><tr><td colspan='2'>[[7b1x]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Uncultured_bacterium Uncultured bacterium]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7B1X OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7B1X FirstGlance]. <br> | ||
</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=7b1x FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7b1x OCA], [https://pdbe.org/7b1x PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7b1x RCSB], [https://www.ebi.ac.uk/pdbsum/7b1x PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7b1x ProSAT]</span></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]] 2.3Å</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=7b1x FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7b1x OCA], [https://pdbe.org/7b1x PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7b1x RCSB], [https://www.ebi.ac.uk/pdbsum/7b1x PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7b1x ProSAT]</span></td></tr> | |||
</table> | </table> | ||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
The gene coding for a novel cold-active esterase PMGL3 was previously obtained from a Siberian permafrost metagenomic DNA library and expressed in Escherichia coli. We elucidated the 3D structure of the enzyme which belongs to the hormone-sensitive lipase (HSL) family. Similar to other bacterial HSLs, PMGL3 shares a canonical alpha/beta hydrolase fold and is presumably a dimer in solution but, in addition to the dimer, it forms a tetrameric structure in a crystal and upon prolonged incubation at 4 degrees C. Detailed analysis demonstrated that the crystal tetramer of PMGL3 has a unique architecture compared to other known tetramers of the bacterial HSLs. To study the role of the specific residues comprising the tetramerization interface of PMGL3, several mutant variants were constructed. Size exclusion chromatography (SEC) analysis of D7N, E47Q, and K67A mutants demonstrated that they still contained a portion of tetrameric form after heat treatment, although its amount was significantly lower in D7N and K67A compared to the wild type. Moreover, the D7N and K67A mutants demonstrated a 40 and 60% increase in the half-life at 40 degrees C in comparison with the wild type protein. K m values of these mutants were similar to that of the wt PMGL3. However, the catalytic constants of the E47Q and K67A mutants were reduced by ~40%. | |||
Structural and Biochemical Characterization of a Cold-Active PMGL3 Esterase with Unusual Oligomeric Structure.,Boyko KM, Kryukova MV, Petrovskaya LE, Kryukova EA, Nikolaeva AY, Korzhenevsky DA, Lomakina GY, Novototskaya-Vlasova KA, Rivkina EM, Dolgikh DA, Kirpichnikov MP, Popov VO Biomolecules. 2021 Jan 5;11(1). pii: biom11010057. doi: 10.3390/biom11010057. PMID:33466452<ref>PMID:33466452</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 7b1x" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Uncultured bacterium]] | |||
[[Category: Boyko KM]] | [[Category: Boyko KM]] | ||
[[Category: Dolgikh DA]] | [[Category: Dolgikh DA]] | ||
Latest revision as of 15:20, 1 February 2024
Crystal structure of cold-active esterase PMGL3 from permafrost metagenomic libraryCrystal structure of cold-active esterase PMGL3 from permafrost metagenomic library
Structural highlights
Publication Abstract from PubMedThe gene coding for a novel cold-active esterase PMGL3 was previously obtained from a Siberian permafrost metagenomic DNA library and expressed in Escherichia coli. We elucidated the 3D structure of the enzyme which belongs to the hormone-sensitive lipase (HSL) family. Similar to other bacterial HSLs, PMGL3 shares a canonical alpha/beta hydrolase fold and is presumably a dimer in solution but, in addition to the dimer, it forms a tetrameric structure in a crystal and upon prolonged incubation at 4 degrees C. Detailed analysis demonstrated that the crystal tetramer of PMGL3 has a unique architecture compared to other known tetramers of the bacterial HSLs. To study the role of the specific residues comprising the tetramerization interface of PMGL3, several mutant variants were constructed. Size exclusion chromatography (SEC) analysis of D7N, E47Q, and K67A mutants demonstrated that they still contained a portion of tetrameric form after heat treatment, although its amount was significantly lower in D7N and K67A compared to the wild type. Moreover, the D7N and K67A mutants demonstrated a 40 and 60% increase in the half-life at 40 degrees C in comparison with the wild type protein. K m values of these mutants were similar to that of the wt PMGL3. However, the catalytic constants of the E47Q and K67A mutants were reduced by ~40%. Structural and Biochemical Characterization of a Cold-Active PMGL3 Esterase with Unusual Oligomeric Structure.,Boyko KM, Kryukova MV, Petrovskaya LE, Kryukova EA, Nikolaeva AY, Korzhenevsky DA, Lomakina GY, Novototskaya-Vlasova KA, Rivkina EM, Dolgikh DA, Kirpichnikov MP, Popov VO Biomolecules. 2021 Jan 5;11(1). pii: biom11010057. doi: 10.3390/biom11010057. PMID:33466452[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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