View source for 8ifj


==Crystal structure of pyrrolysyl-tRNA synthetase from methanogenic archaeon ISO4-G1==
<StructureSection load='8ifj' size='340' side='right'caption='[[8ifj]], [[Resolution|resolution]] 2.78&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[8ifj]] is a 10 chain structure with sequence from [https://en.wikipedia.org/wiki/Methanogenic_archaeon_mixed_culture_ISO4-G1 Methanogenic archaeon mixed culture ISO4-G1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=8IFJ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=8IFJ FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.78&#8491;</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=8ifj FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=8ifj OCA], [https://pdbe.org/8ifj PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=8ifj RCSB], [https://www.ebi.ac.uk/pdbsum/8ifj PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=8ifj ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/A0A126QV54_9EURY A0A126QV54_9EURY] 
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== Publication Abstract from PubMed ==
Pairs of pyrrolysyl-tRNA synthetase (PylRS) and tRNA(Pyl) from Methanosarcina mazei and Methanosarcina barkeri are widely used for site-specific incorporations of non-canonical amino acids into proteins (genetic code expansion). Previously, we achieved full productivity of cell-free protein synthesis for bulky non-canonical amino acids, including N(epsilon)-((((E)-cyclooct-2-en-1-yl)oxy)carbonyl)-L-lysine (TCO*Lys), by using Methanomethylophilus alvus PylRS with structure-based mutations in and around the amino acid binding pocket (first-layer and second-layer mutations, respectively). Recently, the PylRS.tRNA(Pyl) pair from a methanogenic archaeon ISO4-G1 was used for genetic code expansion. In the present study, we determined the crystal structure of the methanogenic archaeon ISO4-G1 PylRS (ISO4-G1 PylRS) and compared it with those of structure-known PylRSs. Based on the ISO4-G1 PylRS structure, we attempted the site-specific incorporation of N(epsilon)-(p-ethynylbenzyloxycarbonyl)-L-lysine (pEtZLys) into proteins, but it was much less efficient than that of TCO*Lys with M. alvus PylRS mutants. Thus, the first-layer mutations (Y125A and M128L) of ISO4-G1 PylRS, with no additional second-layer mutations, increased the protein productivity with pEtZLys up to 57 +/- 8% of that with TCO*Lys at high enzyme concentrations in the cell-free protein synthesis.

Crystal Structure of Pyrrolysyl-tRNA Synthetase from a Methanogenic Archaeon ISO4-G1 and Its Structure-Based Engineering for Highly-Productive Cell-Free Genetic Code Expansion with Non-Canonical Amino Acids.,Yanagisawa T, Seki E, Tanabe H, Fujii Y, Sakamoto K, Yokoyama S Int J Mol Sci. 2023 Mar 26;24(7):6256. doi: 10.3390/ijms24076256. PMID:37047230<ref>PMID:37047230</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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<div class="pdbe-citations 8ifj" style="background-color:#fffaf0;"></div>
== References ==
<references/>
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</StructureSection>
[[Category: Large Structures]]
[[Category: Methanogenic archaeon mixed culture ISO4-G1]]
[[Category: Tanabe H]]
[[Category: Yanagisawa T]]
[[Category: Yokoyama S]]