4k1v: Difference between revisions
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==Crystal structure of delta5-3-ketosteroid isomerase containing Y16F and Y57F mutations== | |||
<StructureSection load='4k1v' size='340' side='right' caption='[[4k1v]], [[Resolution|resolution]] 1.80Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[4k1v]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4K1V OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4K1V FirstGlance]. <br> | |||
</td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4k1u|4k1u]]</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 Pseudomonas putida])</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> | |||
<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=4k1v FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4k1v OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4k1v RCSB], [http://www.ebi.ac.uk/pdbsum/4k1v PDBsum]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
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. | |||
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> | |||
== | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
</div> | |||
==See Also== | |||
*[[Ketosteroid Isomerase|Ketosteroid Isomerase]] | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Pseudomonas putida]] | [[Category: Pseudomonas putida]] | ||
[[Category: Steroid Delta-isomerase]] | [[Category: Steroid Delta-isomerase]] | ||
[[Category: Cha, H J | [[Category: Cha, H J]] | ||
[[Category: Choi, K Y | [[Category: Choi, K Y]] | ||
[[Category: Hong, B H | [[Category: Hong, B H]] | ||
[[Category: Jang, D S | [[Category: Jang, D S]] | ||
[[Category: Kim, Y G | [[Category: Kim, Y G]] | ||
[[Category: Woo, J S | [[Category: Woo, J S]] | ||
[[Category: Allylic isomerization]] | [[Category: Allylic isomerization]] | ||
[[Category: Cystatin-like fold]] | [[Category: Cystatin-like fold]] | ||
