7k0a - Revision history

OCA at 15:15, 18 October 2023

OCA — Wed, 18 Oct 2023 15:15:32 GMT

←Older revision		Revision as of 15:15, 18 October 2023
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	==Puromycin N-acetyltransferase in complex with acetylated puromycin and CoA==		==Puromycin N-acetyltransferase in complex with acetylated puromycin and CoA==
-	<StructureSection load='7k0a' size='340' side='right'caption='[[7k0a]]' scene=''>	+	<StructureSection load='7k0a' size='340' side='right'caption='[[7k0a]], [[Resolution\|resolution]] 2.00Å' 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=7K0A OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7K0A FirstGlance]. <br>	+	<table><tr><td colspan='2'>[[7k0a]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Streptomyces_alboniger Streptomyces alboniger]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7K0A OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7K0A FirstGlance]. <br>
-	</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=7k0a FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7k0a OCA], [https://pdbe.org/7k0a PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7k0a RCSB], [https://www.ebi.ac.uk/pdbsum/7k0a PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7k0a ProSAT]</span></td></tr>	+	</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models\|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution\|Resolution]] 2Å</td></tr>
		+	<tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=COA:COENZYME+A'>COA</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=PEG:DI(HYDROXYETHYL)ETHER'>PEG</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>, <scene name='pdbligand=VQ1:N-Acetylpuromycin'>VQ1</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=7k0a FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7k0a OCA], [https://pdbe.org/7k0a PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7k0a RCSB], [https://www.ebi.ac.uk/pdbsum/7k0a PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7k0a ProSAT]</span></td></tr>
	</table>		</table>
		+	== Function ==
		+	[https://www.uniprot.org/uniprot/PUAC_STRAD PUAC_STRAD] Detoxification of puromycin.
		+	<div style="background-color:#fffaf0;">
		+	== Publication Abstract from PubMed ==
		+	Puromycin and the Streptomyces alboniger-derived puromycin N-acetyltransferase (PAC) enzyme form a commonly used system for selecting stably transfected cultured cells. The crystal structure of PAC has been solved using X-ray crystallography, revealing it to be a member of the GCN5-related N-acetyltransferase (GNAT) family of acetyltransferases. Based on structures in complex with acetyl-CoA or the reaction products CoA and acetylated puromycin, four classes of mutations in and around the catalytic site were designed and tested for activity. Single-residue mutations were identified that displayed a range of enzymatic activities, from complete ablation to enhanced activity relative to wild-type (WT) PAC. Cell pools of stably transfected HEK293 cells derived using two PAC mutants with attenuated activity, Y30F and A142D, were found to secrete up to three-fold higher levels of a soluble, recombinant target protein than corresponding pools derived with the WT enzyme. A third mutant, Y171F, appeared to stabilise the intracellular turnover of PAC, resulting in an apparent loss of selection stringency. Our results indicate that the structure-guided manipulation of PAC function can be utilised to enhance selection stringency for the derivation of mammalian cell lines secreting elevated levels of recombinant proteins.
		+
		+	Structure-guided selection of puromycin N-acetyltransferase mutants with enhanced selection stringency for deriving mammalian cell lines expressing recombinant proteins.,Caputo AT, Eder OM, Bereznakova H, Pothuis H, Ardevol A, Newman J, Nuttall S, Peat TS, Adams TE Sci Rep. 2021 Mar 4;11(1):5247. doi: 10.1038/s41598-021-84551-9. PMID:33664348<ref>PMID:33664348</ref>
		+
		+	From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
		+	</div>
		+	<div class="pdbe-citations 7k0a" style="background-color:#fffaf0;"></div>
		+	== References ==
		+	<references/>
	__TOC__		__TOC__
	</StructureSection>		</StructureSection>
	[[Category: Large Structures]]		[[Category: Large Structures]]
		+	[[Category: Streptomyces alboniger]]
	[[Category: Adams TE]]		[[Category: Adams TE]]
	[[Category: Caputo AT]]		[[Category: Caputo AT]]
	[[Category: Newman J]]		[[Category: Newman J]]
	[[Category: Peat TS]]		[[Category: Peat TS]]

OCA at 06:48, 17 March 2021

OCA — Wed, 17 Mar 2021 06:48:02 GMT

←Older revision		Revision as of 06:48, 17 March 2021
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-	'''Unreleased structure'''

-	~~The entry~~ 7k0a ~~is ON HOLD until Paper Publication~~	+	==Puromycin N-acetyltransferase in complex with acetylated puromycin and CoA==
-		+	<StructureSection load='7k0a' size='340' side='right'caption='[[7k0a]]' scene=''>
-	~~Authors~~: ~~Peat~~, T.S., ~~Caputo~~, A.T., ~~Newman, J~~., ~~Adams, T~~.E.	+	== Structural highlights ==
-		+	<table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7K0A OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7K0A FirstGlance]. <br>
-	~~Description: Puromycin N-acetyltransferase in complex with acetylated puromycin and CoA~~	+	</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=7k0a FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7k0a OCA], [https://pdbe.org/7k0a PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7k0a RCSB], [https://www.ebi.ac.uk/pdbsum/7k0a PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7k0a ProSAT]</span></td></tr>
-	[[Category: ~~Unreleased~~ Structures]]	+	</table>
-	[[Category: ~~Peat, T.S~~]]	+	__TOC__
-	[[Category: Caputo~~, A.T~~]]	+	</StructureSection>
-	[[Category: Newman, J]]	+	[[Category: Large Structures]]
-	[[Category: ~~Adams, T.E~~]]	+	[[Category: Adams TE]]
		+	[[Category: Caputo AT]]
		+	[[Category: Newman J]]
		+	[[Category: Peat TS]]

OCA at 07:02, 16 September 2020

OCA — Wed, 16 Sep 2020 07:02:51 GMT

←Older revision		Revision as of 07:02, 16 September 2020
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	'''Unreleased structure'''		'''Unreleased structure'''

-	The entry 7k0a is ON HOLD	+	The entry 7k0a is ON HOLD until Paper Publication

	Authors: Peat, T.S., Caputo, A.T., Newman, J., Adams, T.E.		Authors: Peat, T.S., Caputo, A.T., Newman, J., Adams, T.E.

OCA: Protected "7k0a" [edit=sysop:move=sysop]

OCA — Wed, 09 Sep 2020 07:49:04 GMT

Protected "7k0a" [edit=sysop:move=sysop]

←Older revision

Revision as of 07:49, 9 September 2020

OCA: New page: '''Unreleased structure''' The entry 7k0a is ON HOLD Authors: Peat, T.S., Caputo, A.T., Newman, J., Adams, T.E. Description: Puromycin N-acetyltransferase in complex with acetylated pu...

OCA — Wed, 09 Sep 2020 07:49:03 GMT

New page: '''Unreleased structure''' The entry 7k0a is ON HOLD Authors: Peat, T.S., Caputo, A.T., Newman, J., Adams, T.E. Description: Puromycin N-acetyltransferase in complex with acetylated pu...

New page

'''Unreleased structure'''

The entry 7k0a is ON HOLD

Authors: Peat, T.S., Caputo, A.T., Newman, J., Adams, T.E.

Description: Puromycin N-acetyltransferase in complex with acetylated puromycin and CoA
[[Category: Unreleased Structures]]
[[Category: Peat, T.S]]
[[Category: Caputo, A.T]]
[[Category: Newman, J]]
[[Category: Adams, T.E]]