000818274 001__ 818274
000818274 005__ 20210129224126.0
000818274 0247_ $$2doi$$a10.1039/C5SC04574F
000818274 0247_ $$2ISSN$$a2041-6520
000818274 0247_ $$2ISSN$$a2041-6539
000818274 0247_ $$2Handle$$a2128/12295
000818274 0247_ $$2WOS$$aWOS:000378715000066
000818274 0247_ $$2altmetric$$aaltmetric:9576342
000818274 0247_ $$2pmid$$apmid:30155096
000818274 037__ $$aFZJ-2016-04748
000818274 041__ $$aEnglish
000818274 082__ $$a540
000818274 1001_ $$0P:(DE-Juel1)136932$$aDick, Markus$$b0
000818274 245__ $$aMechanism-based inhibition of an aldolase at high concentrations of its natural substrate acetaldehyde: structural insights and protective strategies
000818274 260__ $$aCambridge$$bRSC$$c2016
000818274 3367_ $$2DRIVER$$aarticle
000818274 3367_ $$2DataCite$$aOutput Types/Journal article
000818274 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article$$bjournal$$mjournal$$s1485183783_18866
000818274 3367_ $$2BibTeX$$aARTICLE
000818274 3367_ $$2ORCID$$aJOURNAL_ARTICLE
000818274 3367_ $$00$$2EndNote$$aJournal Article
000818274 520__ $$a2-Deoxy-D-ribose-5-phosphate aldolase (DERA) is used in organic synthesis for the enantioselective reaction between acetaldehyde and a broad range of other aldehydes as acceptor molecules. Nevertheless, its application is hampered by a poor tolerance towards high concentrations of acetaldehyde, its natural substrate. While numerous studies have been performed searching for new, more acetaldehyde-resistant DERAs, the mechanism underlying this deactivation process has remained elusive. By using NMR spectroscopy on both the protein and the small-molecule scale, we could show that a reaction product binds to the inner part of the enzyme, and that this effect can be partly reversed via heating. The crystal structure of DERA before and after acetaldehyde incubation was determined at high resolution, revealing a covalently bound reaction product bridging the catalytically active lysine (K167) to a nearby cysteine (C47) in the deactivated enzyme. A reaction mechanism is proposed where crotonaldehyde as the aldol product of two acetaldehyde molecules after water elimination forms a Schiff base with the lysine side chain, followed by Michael addition of the cysteine thiol group to the Cβ atom of the inhibitor. In support of this mechanism, direct incubation of DERA with crotonaldehyde results in a more than 100-fold stronger inhibition, compared to acetaldehyde, whereas mutation of C47 gives rise to a fully acetaldehyde-resistant DERA. Thus this variant appears perfectly suited for synthetic applications. A similar diagnostic and preventive strategy should be applicable to other biocatalysts suffering from mechanism-based inhibition by a reactive substrate, a condition that may be more common than currently appreciated in biotechnology.
000818274 536__ $$0G:(DE-HGF)POF3-553$$a553 - Physical Basis of Diseases (POF3-553)$$cPOF3-553$$fPOF III$$x0
000818274 536__ $$0G:(DE-HGF)POF3-581$$a581 - Biotechnology (POF3-581)$$cPOF3-581$$fPOF III$$x1
000818274 588__ $$aDataset connected to CrossRef
000818274 7001_ $$0P:(DE-Juel1)132001$$aHartmann, Rudolf$$b1
000818274 7001_ $$0P:(DE-Juel1)131988$$aWeiergräber, Oliver H.$$b2
000818274 7001_ $$0P:(DE-Juel1)156323$$aBisterfeld, Carolin$$b3
000818274 7001_ $$0P:(DE-Juel1)128890$$aClassen, Thomas$$b4
000818274 7001_ $$0P:(DE-Juel1)132019$$aSchwarten, Melanie$$b5
000818274 7001_ $$0P:(DE-Juel1)144510$$aNeudecker, Philipp$$b6
000818274 7001_ $$0P:(DE-Juel1)132029$$aWillbold, Dieter$$b7
000818274 7001_ $$0P:(DE-Juel1)128906$$aPietruszka, Jörg$$b8$$eCorresponding author
000818274 773__ $$0PERI:(DE-600)2559110-1$$a10.1039/C5SC04574F$$gVol. 7, no. 7, p. 4492 - 4502$$n7$$p4492 - 4502$$tChemical science$$v7$$x2041-6539$$y2016
000818274 8564_ $$uhttps://juser.fz-juelich.de/record/818274/files/Mechanism-based%20inhibition%20of%20an%20aldolase%20at%20high%20concentrations%20of%20its%20natural%20substrate%20acetaldehyde%3A%20structural%20insights%20and%20protective%20strategies.pdf$$yOpenAccess
000818274 8564_ $$uhttps://juser.fz-juelich.de/record/818274/files/Mechanism-based%20inhibition%20of%20an%20aldolase%20at%20high%20concentrations%20of%20its%20natural%20substrate%20acetaldehyde%3A%20structural%20insights%20and%20protective%20strategies.gif?subformat=icon$$xicon$$yOpenAccess
000818274 8564_ $$uhttps://juser.fz-juelich.de/record/818274/files/Mechanism-based%20inhibition%20of%20an%20aldolase%20at%20high%20concentrations%20of%20its%20natural%20substrate%20acetaldehyde%3A%20structural%20insights%20and%20protective%20strategies.jpg?subformat=icon-1440$$xicon-1440$$yOpenAccess
000818274 8564_ $$uhttps://juser.fz-juelich.de/record/818274/files/Mechanism-based%20inhibition%20of%20an%20aldolase%20at%20high%20concentrations%20of%20its%20natural%20substrate%20acetaldehyde%3A%20structural%20insights%20and%20protective%20strategies.jpg?subformat=icon-180$$xicon-180$$yOpenAccess
000818274 8564_ $$uhttps://juser.fz-juelich.de/record/818274/files/Mechanism-based%20inhibition%20of%20an%20aldolase%20at%20high%20concentrations%20of%20its%20natural%20substrate%20acetaldehyde%3A%20structural%20insights%20and%20protective%20strategies.jpg?subformat=icon-640$$xicon-640$$yOpenAccess
000818274 8564_ $$uhttps://juser.fz-juelich.de/record/818274/files/Mechanism-based%20inhibition%20of%20an%20aldolase%20at%20high%20concentrations%20of%20its%20natural%20substrate%20acetaldehyde%3A%20structural%20insights%20and%20protective%20strategies.pdf?subformat=pdfa$$xpdfa$$yOpenAccess
000818274 909CO $$ooai:juser.fz-juelich.de:818274$$pdnbdelivery$$pVDB$$pdriver$$popen_access$$popenaire
000818274 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)132001$$aForschungszentrum Jülich$$b1$$kFZJ
000818274 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)131988$$aForschungszentrum Jülich$$b2$$kFZJ
000818274 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)156323$$aForschungszentrum Jülich$$b3$$kFZJ
000818274 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)128890$$aForschungszentrum Jülich$$b4$$kFZJ
000818274 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)132019$$aForschungszentrum Jülich$$b5$$kFZJ
000818274 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)144510$$aForschungszentrum Jülich$$b6$$kFZJ
000818274 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)132029$$aForschungszentrum Jülich$$b7$$kFZJ
000818274 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)128906$$aForschungszentrum Jülich$$b8$$kFZJ
000818274 9131_ $$0G:(DE-HGF)POF3-553$$1G:(DE-HGF)POF3-550$$2G:(DE-HGF)POF3-500$$3G:(DE-HGF)POF3$$4G:(DE-HGF)POF$$aDE-HGF$$bKey Technologies$$lBioSoft – Fundamentals for future Technologies in the fields of Soft Matter and Life Sciences$$vPhysical Basis of Diseases$$x0
000818274 9131_ $$0G:(DE-HGF)POF3-581$$1G:(DE-HGF)POF3-580$$2G:(DE-HGF)POF3-500$$3G:(DE-HGF)POF3$$4G:(DE-HGF)POF$$aDE-HGF$$bKey Technologies$$lKey Technologies for the Bioeconomy$$vBiotechnology$$x1
000818274 9141_ $$y2016
000818274 915__ $$0LIC:(DE-HGF)CCBY3$$2HGFVOC$$aCreative Commons Attribution CC BY 3.0
000818274 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS
000818274 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR$$bCHEM SCI : 2015
000818274 915__ $$0StatID:(DE-HGF)9905$$2StatID$$aIF >= 5$$bCHEM SCI : 2015
000818274 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection
000818274 915__ $$0StatID:(DE-HGF)0110$$2StatID$$aWoS$$bScience Citation Index
000818274 915__ $$0StatID:(DE-HGF)0111$$2StatID$$aWoS$$bScience Citation Index Expanded
000818274 915__ $$0StatID:(DE-HGF)0510$$2StatID$$aOpenAccess
000818274 915__ $$0StatID:(DE-HGF)1150$$2StatID$$aDBCoverage$$bCurrent Contents - Physical, Chemical and Earth Sciences
000818274 915__ $$0StatID:(DE-HGF)0310$$2StatID$$aDBCoverage$$bNCBI Molecular Biology Database
000818274 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline
000818274 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bThomson Reuters Master Journal List
000818274 920__ $$lyes
000818274 9201_ $$0I:(DE-Juel1)ICS-6-20110106$$kICS-6$$lStrukturbiochemie $$x0
000818274 9201_ $$0I:(DE-Juel1)IBOC-20090406$$kIBOC$$lInstitut für Bioorganische Chemie (HHUD)$$x1
000818274 9201_ $$0I:(DE-Juel1)IBG-1-20101118$$kIBG-1$$lBiotechnologie$$x2
000818274 9801_ $$aFullTexts
000818274 980__ $$ajournal
000818274 980__ $$aVDB
000818274 980__ $$aI:(DE-Juel1)ICS-6-20110106
000818274 980__ $$aI:(DE-Juel1)IBOC-20090406
000818274 980__ $$aI:(DE-Juel1)IBG-1-20101118
000818274 980__ $$aUNRESTRICTED
000818274 981__ $$aI:(DE-Juel1)IBI-7-20200312