6frh: Difference between revisions
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==Crystal structure of Ssp DnaB Mini-Intein variant M86== | |||
<StructureSection load='6frh' size='340' side='right' caption='[[6frh]], [[Resolution|resolution]] 2.03Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[6frh]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Syny3 Syny3]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6FRH OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6FRH FirstGlance]. <br> | |||
</td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[6fre|6fre]], [[6frg|6frg]]</td></tr> | |||
<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">dnaB, slr0833 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1111708 SYNY3])</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=6frh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6frh OCA], [http://pdbe.org/6frh PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6frh RCSB], [http://www.ebi.ac.uk/pdbsum/6frh PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6frh ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[[http://www.uniprot.org/uniprot/DNAB_SYNY3 DNAB_SYNY3]] Participates in initiation and elongation during chromosome replication; it exhibits DNA-dependent ATPase activity (By similarity). | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Inteins remove themselves from a precursor protein by protein splicing. Due to the concomitant structural changes of the host protein, this self-processing reaction has enabled many applications in protein biotechnology and chemical biology. We show that the evolved M86 mutant of the Ssp DnaB intein displays a significantly improved tolerance towards non-native amino acids at the N-terminally flanking (-1) extein position compared to the parent intein, in the form of both an artificially trans-splicing split intein and the cis-splicing mini-intein. Surprisingly, side chains with increased steric bulk compared to the native Gly(-1) residue, including d-amino acids, were found to compensate for the essential block B histidine in His73Ala mutants in the initial N-S acyl shift of the protein splicing pathway. In the case of the M86 intein, large (-1) side chains can even rescue protein splicing activity as a whole. With the comparison of three crystal structures, namely of the M86 intein as well as of its Gly(-1)Phe and Gly(-1)Phe/His73Ala mutants, our data supports a model in which the intein's active site can exert a strain by varying mechanisms on the different angles of the scissile bond at the extein-intein junction to effect a ground-state destabilization. The compensatory mechanism of the block B histidine is the first example for the direct functional role of an extein residue in protein splicing. It sheds new light on the extein-intein interplay and on possible consequences of their co-evolution as well as on the laboratory engineering of improved inteins. | |||
A functional interplay between intein and extein sequences in protein splicing compensates for the essential block B histidine.,Friedel K, Popp MA, Matern JCJ, Gazdag EM, Thiel IV, Volkmann G, Blankenfeldt W, Mootz HD Chem Sci. 2018 Oct 3;10(1):239-251. doi: 10.1039/c8sc01074a. eCollection 2019 Jan, 7. PMID:30713635<ref>PMID:30713635</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 6frh" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Syny3]] | |||
[[Category: Blankenfeldt, W]] | |||
[[Category: Gazdag, M E]] | |||
[[Category: Matern, J J.C]] | |||
[[Category: Mootz, H D]] | |||
[[Category: Popp, M A]] | |||
[[Category: Hint domain fold]] | |||
[[Category: Intein]] | |||
[[Category: Pre-splicing]] | |||
[[Category: Protein splicing]] | |||
[[Category: Splicing]] |