7a3m: Difference between revisions
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==Synergistic stabilization of a double mutant in CI2 from an in-cell library screen== | ==Synergistic stabilization of a double mutant in CI2 from an in-cell library screen== | ||
<StructureSection load='7a3m' size='340' side='right'caption='[[7a3m]]' scene=''> | <StructureSection load='7a3m' size='340' side='right'caption='[[7a3m]], [[Resolution|resolution]] 1.01Å' 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=7A3M OCA]. For a <b>guided tour on the structure components</b> use [ | <table><tr><td colspan='2'>[[7a3m]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Barley Barley]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7A3M OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7A3M FirstGlance]. <br> | ||
</td></tr><tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[ | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat"><div style='overflow: auto; max-height: 3em;'>[[7a1h|7a1h]], [[7a2c|7a2c]], [[7a2e|7a2e]]</div></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=7a3m FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7a3m OCA], [https://pdbe.org/7a3m PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7a3m RCSB], [https://www.ebi.ac.uk/pdbsum/7a3m PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7a3m ProSAT]</span></td></tr> | |||
</table> | </table> | ||
== Function == | |||
[[https://www.uniprot.org/uniprot/ICI2_HORVU ICI2_HORVU]] Inhibits both subtilisin and chymotrypsin. | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Most single point mutations destabilize folded proteins. Mutations that stabilize a protein typically only have a small effect and multiple mutations are often needed to substantially increase the stability. Multiple point mutations may act synergistically on the stability, and it is often not straightforward to predict their combined effect from the individual contributions. Here, we have applied an efficient in-cell assay in E. coli to select variants of the barley chymotrypsin inhibitor 2 with increased stability. We find two variants that are more than 3.8 kJ mol(-1) more stable than the wild-type. In one case, the increased stability is the effect of the single substitution D55G. The other case is a double mutant, L49I/I57V, which is 5.1 kJ mol(-1) more stable than the sum of the effects of the individual mutations. In addition to demonstrating the strength of our selection system for finding stabilizing mutations, our work also demonstrate how subtle conformational effects may modulate stability. | |||
Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli.,Hamborg L, Granata D, Olsen JG, Roche JV, Pedersen LE, Nielsen AT, Lindorff-Larsen K, Teilum K Commun Biol. 2021 Aug 18;4(1):980. doi: 10.1038/s42003-021-02490-7. PMID:34408246<ref>PMID:34408246</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 7a3m" style="background-color:#fffaf0;"></div> | |||
==See Also== | |||
*[[Chymotrypsin inhibitor 3D structures|Chymotrypsin inhibitor 3D structures]] | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Barley]] | |||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Hamborg L]] | [[Category: Hamborg, L]] | ||
[[Category: Olsen | [[Category: Olsen, J G]] | ||
[[Category: Roche | [[Category: Roche, J V]] | ||
[[Category: Teilum K]] | [[Category: Teilum, K]] | ||
[[Category: Chymotrypsin inhibitor ci-2]] | |||
[[Category: Mutant]] | |||
[[Category: Protein binding]] | |||
Revision as of 11:57, 29 September 2021
Synergistic stabilization of a double mutant in CI2 from an in-cell library screenSynergistic stabilization of a double mutant in CI2 from an in-cell library screen
Structural highlights
Function[ICI2_HORVU] Inhibits both subtilisin and chymotrypsin. Publication Abstract from PubMedMost single point mutations destabilize folded proteins. Mutations that stabilize a protein typically only have a small effect and multiple mutations are often needed to substantially increase the stability. Multiple point mutations may act synergistically on the stability, and it is often not straightforward to predict their combined effect from the individual contributions. Here, we have applied an efficient in-cell assay in E. coli to select variants of the barley chymotrypsin inhibitor 2 with increased stability. We find two variants that are more than 3.8 kJ mol(-1) more stable than the wild-type. In one case, the increased stability is the effect of the single substitution D55G. The other case is a double mutant, L49I/I57V, which is 5.1 kJ mol(-1) more stable than the sum of the effects of the individual mutations. In addition to demonstrating the strength of our selection system for finding stabilizing mutations, our work also demonstrate how subtle conformational effects may modulate stability. Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli.,Hamborg L, Granata D, Olsen JG, Roche JV, Pedersen LE, Nielsen AT, Lindorff-Larsen K, Teilum K Commun Biol. 2021 Aug 18;4(1):980. doi: 10.1038/s42003-021-02490-7. PMID:34408246[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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