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| | ==Crystal structure of delta5-3-ketosteroid isomerase containing Y16F and Y32F mutations== | | ==Crystal structure of delta5-3-ketosteroid isomerase containing Y16F and Y32F mutations== |
| - | <StructureSection load='4k1u' size='340' side='right' caption='[[4k1u]], [[Resolution|resolution]] 2.00Å' scene=''> | + | <StructureSection load='4k1u' size='340' side='right'caption='[[4k1u]], [[Resolution|resolution]] 2.00Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4k1u]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/"bacillus_fluorescens_putidus"_flugge_1886 "bacillus fluorescens putidus" flugge 1886]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4K1U OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4K1U FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4k1u]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4K1U OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4K1U FirstGlance]. <br> |
| - | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[1opy|1opy]], [[4k1v|4k1v]]</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]] 2Å</td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">ksi ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=303 "Bacillus fluorescens putidus" Flugge 1886])</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=4k1u FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4k1u OCA], [https://pdbe.org/4k1u PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4k1u RCSB], [https://www.ebi.ac.uk/pdbsum/4k1u PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4k1u ProSAT]</span></td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Steroid_Delta-isomerase Steroid Delta-isomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.3.3.1 5.3.3.1] </span></td></tr>
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| - | <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=4k1u FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4k1u OCA], [http://pdbe.org/4k1u PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4k1u RCSB], [http://www.ebi.ac.uk/pdbsum/4k1u PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4k1u ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| - | <div style="background-color:#fffaf0;">
| + | == Function == |
| - | == Publication Abstract from PubMed == | + | [https://www.uniprot.org/uniprot/SDIS_PSEPU SDIS_PSEPU] |
| - | Proteins have evolved to compensate for detrimental mutations. However, compensatory mechanisms for protein defects are not well understood. Using ketosteroid isomerase (KSI), we investigated how second-site mutations could recover defective mutant function and stability. Previous results revealed that the Y30F mutation rescued the Y14F, Y55F and Y14F/Y55F mutants by increasing the catalytic activity by 23-, 3- and 1.3-fold, respectively, and the Y55F mutant by increasing the stability by 3.3 kcal/mol. To better understand these observations, we systematically investigated detailed structural and thermodynamic effects of the Y30F mutation on these mutants. Crystal structures of the Y14F/Y30F and Y14F/Y55F mutants were solved at 2.0 and 1.8 previoulsy solved structures of wild-type and other mutant KSIs. Structural analyses revealed that the Y30F mutation partially restored the active-site cleft of these mutant KSIs. The Y30F mutation also increased Y14F and Y14F/Y55F mutant stability by 3.2 and 4.3 kcal/mol, respectively, and the melting temperatures of the Y14F, Y55F and Y14F/Y55F mutants by 6.4 degrees C, 5.1 degrees C and 10.0 degrees C, respectively. Compensatory effects of the Y30F mutation on stability might be due to improved hydrophobic interactions because removal of a hydroxyl group from Tyr30 induced local compaction by neighboring residue movement and enhanced interactions with surrounding hydrophobic residues in the active site. Taken together, our results suggest that perturbed active-site geometry recovery and favorable hydrophobic interactions mediate the role of Y30F as a secondsite suppressor.
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| - | Rescue of deleterious mutations by the compensatory Y30F mutation in ketosteroid isomerase.,Cha HJ, Jang do S, Kim YG, Hong BH, Woo JS, Kim KT, Choi KY Mol Cells. 2013 Jul;36(1):39-46. doi: 10.1007/s10059-013-0013-1. Epub 2013 Jun 3. PMID:23740430<ref>PMID:23740430</ref>
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| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| - | </div>
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| - | <div class="pdbe-citations 4k1u" style="background-color:#fffaf0;"></div>
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| | ==See Also== | | ==See Also== |
| | *[[Ketosteroid Isomerase|Ketosteroid Isomerase]] | | *[[Ketosteroid Isomerase|Ketosteroid Isomerase]] |
| - | == References == | |
| - | <references/> | |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Bacillus fluorescens putidus flugge 1886]] | + | [[Category: Large Structures]] |
| - | [[Category: Steroid Delta-isomerase]] | + | [[Category: Pseudomonas putida]] |
| - | [[Category: Cha, H J]] | + | [[Category: Cha HJ]] |
| - | [[Category: Choi, K Y]] | + | [[Category: Choi KY]] |
| - | [[Category: Hong, B H]] | + | [[Category: Hong BH]] |
| - | [[Category: Jang, D S]] | + | [[Category: Jang DS]] |
| - | [[Category: Kim, Y G]] | + | [[Category: Kim YG]] |
| - | [[Category: Woo, J S]] | + | [[Category: Woo JS]] |
| - | [[Category: Allylic isomerization]]
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| - | [[Category: Cystatin-like fold]]
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| - | [[Category: Cytosol]]
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| - | [[Category: Delta5-3ketosteroid]]
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| - | [[Category: Isomerase]]
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