7xws: Difference between revisions
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==Crystal structure of Wild Type Cypovirus Polyhedra produced by cell-free protein synthesis with small volume== | |||
<StructureSection load='7xws' size='340' side='right'caption='[[7xws]], [[Resolution|resolution]] 1.95Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[7xws]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Bombyx_mori_cypovirus_1 Bombyx mori cypovirus 1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7XWS OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7XWS 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]] 1.95Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACE:ACETYL+GROUP'>ACE</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene></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=7xws FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7xws OCA], [https://pdbe.org/7xws PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7xws RCSB], [https://www.ebi.ac.uk/pdbsum/7xws PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7xws ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/PYHD_CPVBM PYHD_CPVBM] Major component of the virus occlusion bodies, which are large proteinaceous structures (polyhedra), that protect the virus from the outside environment for extended periods until they are ingested by insect larvae. | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
In-cell protein crystallization (ICPC) has been investigated as a technique to support the advancement of structural biology because it does not require protein purification and a complicated crystallization process. However, only a few protein structures have been reported because these crystals formed incidentally in living cells and are insufficient in size and quality for structure analysis. Here, we have developed a cell-free protein crystallization (CFPC) method, which involves direct protein crystallization using cell-free protein synthesis. We have succeeded in crystallization and structure determination of nano-sized polyhedra crystal (PhC) at a high resolution of 1.80 A. Furthermore, nanocrystals were synthesized at a reaction scale of only 20 muL using the dialysis method, enabling structural analysis at a resolution of 1.95 A. To further demonstrate the potential of CFPC, we attempted to determine the structure of crystalline inclusion protein A (CipA), whose structure had not yet been determined. We added chemical reagents as a twinning inhibitor to the CFPC solution, which enabled us to determine the structure of CipA at 2.11 A resolution. This technology greatly expands the high-throughput structure determination method of unstable, low-yield, fusion, and substrate-biding proteins that have been difficult to analyze with conventional methods. | |||
Cell-free protein crystallization for nanocrystal structure determination.,Abe S, Tanaka J, Kojima M, Kanamaru S, Hirata K, Yamashita K, Kobayashi A, Ueno T Sci Rep. 2022 Oct 3;12(1):16031. doi: 10.1038/s41598-022-19681-9. PMID:36192567<ref>PMID:36192567</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
[[Category: | <div class="pdbe-citations 7xws" style="background-color:#fffaf0;"></div> | ||
[[Category: | |||
[[Category: | ==See Also== | ||
[[Category: | *[[Polyhedrin|Polyhedrin]] | ||
[[Category: | == References == | ||
[[Category: Ueno | <references/> | ||
__TOC__ | |||
</StructureSection> | |||
[[Category: Bombyx mori cypovirus 1]] | |||
[[Category: Large Structures]] | |||
[[Category: Abe S]] | |||
[[Category: Hirata K]] | |||
[[Category: Kojima M]] | |||
[[Category: Tanaka J]] | |||
[[Category: Ueno T]] | |||
[[Category: Yamashita K]] | |||
Latest revision as of 21:02, 29 November 2023
Crystal structure of Wild Type Cypovirus Polyhedra produced by cell-free protein synthesis with small volumeCrystal structure of Wild Type Cypovirus Polyhedra produced by cell-free protein synthesis with small volume
Structural highlights
FunctionPYHD_CPVBM Major component of the virus occlusion bodies, which are large proteinaceous structures (polyhedra), that protect the virus from the outside environment for extended periods until they are ingested by insect larvae. Publication Abstract from PubMedIn-cell protein crystallization (ICPC) has been investigated as a technique to support the advancement of structural biology because it does not require protein purification and a complicated crystallization process. However, only a few protein structures have been reported because these crystals formed incidentally in living cells and are insufficient in size and quality for structure analysis. Here, we have developed a cell-free protein crystallization (CFPC) method, which involves direct protein crystallization using cell-free protein synthesis. We have succeeded in crystallization and structure determination of nano-sized polyhedra crystal (PhC) at a high resolution of 1.80 A. Furthermore, nanocrystals were synthesized at a reaction scale of only 20 muL using the dialysis method, enabling structural analysis at a resolution of 1.95 A. To further demonstrate the potential of CFPC, we attempted to determine the structure of crystalline inclusion protein A (CipA), whose structure had not yet been determined. We added chemical reagents as a twinning inhibitor to the CFPC solution, which enabled us to determine the structure of CipA at 2.11 A resolution. This technology greatly expands the high-throughput structure determination method of unstable, low-yield, fusion, and substrate-biding proteins that have been difficult to analyze with conventional methods. Cell-free protein crystallization for nanocrystal structure determination.,Abe S, Tanaka J, Kojima M, Kanamaru S, Hirata K, Yamashita K, Kobayashi A, Ueno T Sci Rep. 2022 Oct 3;12(1):16031. doi: 10.1038/s41598-022-19681-9. PMID:36192567[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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