Crystal structure of a thermostable old yellow enzyme from Thermus scotoductus SA-01

 

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dc.contributor.author Opperman, Diederik J
dc.contributor.author Sewell, Bryan T
dc.contributor.author Litthauer, Derek
dc.contributor.author Isupov, Mikhail N
dc.contributor.author Littlechild, Jennifer A
dc.contributor.author Heerden, Esta van
dc.date.accessioned 2016-08-31T10:54:25Z
dc.date.available 2016-08-31T10:54:25Z
dc.date.issued 2010
dc.identifier http://dx.doi.org/10.1016/j.bbrc.2010.02.011
dc.identifier.citation Opperman, D. J., Sewell, B. T., Litthauer, D., Isupov, M. N., Littlechild, J. A., & van Heerden, E. (2010). Crystal structure of a thermostable old yellow enzyme from Thermus scotoductus SA-01. Biochemical and biophysical research communications, 393(3), 426-431. en_ZA
dc.identifier.issn 0006-291X en_ZA
dc.identifier.uri http://hdl.handle.net/11427/21606
dc.identifier.uri http://www.sciencedirect.com/science/article/pii/S0006291X10002111
dc.description.abstract Recent characterization of the chromate reductase (CrS) from the thermophile Thermus scotoductus SA-01 revealed this enzyme to be related to the Old Yellow Enzyme (OYE) family. Here, we report the structure of a thermostable OYE homolog in its holoform at 2.2 Å as well as its complex with p-hydroxybenzaldehyde (pHBA). The enzyme crystallized as octamers with the monomers showing a classical TIM barrel fold which upon dimerization yields the biologically active form of the protein. A sulfate ion is bound above the si-side of the non-covalently bound FMN cofactor in the oxidized solved structure but is displaced upon pHBA binding. The active-site architecture is highly conserved as with other members of this enzyme family. The pHBA in the CrS complex is positioned by hydrogen bonding to the two conserved catalytic-site histidines. The most prominent structural difference between CrS and other OYE homologs is the size of the “capping domain”. Thermostabilization of the enzyme is achieved in part through increased proline content within loops and turns as well as increased intersubunit interactions through hydrogen bonding and complex salt bridge networks. CrS is able to reduce the C C bonds of α,β-unsaturated carbonyl compounds with a preference towards cyclic substrates however no activity was observed towards β-substituted substrates. Mutational studies have confirmed the role of Tyr177 as the proposed proton donor although reduction could still occur at a reduced rate when this residue was mutated to phenylalanine. en_ZA
dc.language eng en_ZA
dc.publisher Elsevier en_ZA
dc.source Biochemical and Biophysical Research Communications en_ZA
dc.source.uri http://www.journals.elsevier.com/biochemical-and-biophysical-research-communications/
dc.subject.other Old Yellow Enzyme
dc.subject.other Flavoprotein
dc.subject.other Thermus scotoductus
dc.subject.other Thermostability
dc.subject.other Enoate reduction
dc.title Crystal structure of a thermostable old yellow enzyme from Thermus scotoductus SA-01 en_ZA
dc.type Journal Article en_ZA
dc.date.updated 2016-08-31T10:53:07Z
uct.type.publication Research en_ZA
uct.type.resource Article en_ZA
dc.publisher.institution University of Cape Town
uct.type.filetype Text
uct.type.filetype Image
dc.identifier.apacitation Opperman, D. J., Sewell, B. T., Litthauer, D., Isupov, M. N., Littlechild, J. A., & Heerden, E. v. (2010). Crystal structure of a thermostable old yellow enzyme from Thermus scotoductus SA-01. <i>Biochemical and Biophysical Research Communications</i>, http://hdl.handle.net/11427/21606 en_ZA
dc.identifier.chicagocitation Opperman, Diederik J, Bryan T Sewell, Derek Litthauer, Mikhail N Isupov, Jennifer A Littlechild, and Esta van Heerden "Crystal structure of a thermostable old yellow enzyme from Thermus scotoductus SA-01." <i>Biochemical and Biophysical Research Communications</i> (2010) http://hdl.handle.net/11427/21606 en_ZA
dc.identifier.vancouvercitation Opperman DJ, Sewell BT, Litthauer D, Isupov MN, Littlechild JA, Heerden Ev. Crystal structure of a thermostable old yellow enzyme from Thermus scotoductus SA-01. Biochemical and Biophysical Research Communications. 2010; http://hdl.handle.net/11427/21606. en_ZA
dc.identifier.ris TY - Journal Article AU - Opperman, Diederik J AU - Sewell, Bryan T AU - Litthauer, Derek AU - Isupov, Mikhail N AU - Littlechild, Jennifer A AU - Heerden, Esta van AB - Recent characterization of the chromate reductase (CrS) from the thermophile Thermus scotoductus SA-01 revealed this enzyme to be related to the Old Yellow Enzyme (OYE) family. Here, we report the structure of a thermostable OYE homolog in its holoform at 2.2 Å as well as its complex with p-hydroxybenzaldehyde (pHBA). The enzyme crystallized as octamers with the monomers showing a classical TIM barrel fold which upon dimerization yields the biologically active form of the protein. A sulfate ion is bound above the si-side of the non-covalently bound FMN cofactor in the oxidized solved structure but is displaced upon pHBA binding. The active-site architecture is highly conserved as with other members of this enzyme family. The pHBA in the CrS complex is positioned by hydrogen bonding to the two conserved catalytic-site histidines. The most prominent structural difference between CrS and other OYE homologs is the size of the “capping domain”. Thermostabilization of the enzyme is achieved in part through increased proline content within loops and turns as well as increased intersubunit interactions through hydrogen bonding and complex salt bridge networks. CrS is able to reduce the C C bonds of α,β-unsaturated carbonyl compounds with a preference towards cyclic substrates however no activity was observed towards β-substituted substrates. Mutational studies have confirmed the role of Tyr177 as the proposed proton donor although reduction could still occur at a reduced rate when this residue was mutated to phenylalanine. DA - 2010 DB - OpenUCT DP - University of Cape Town J1 - Biochemical and Biophysical Research Communications LK - https://open.uct.ac.za PB - University of Cape Town PY - 2010 SM - 0006-291X T1 - Crystal structure of a thermostable old yellow enzyme from Thermus scotoductus SA-01 TI - Crystal structure of a thermostable old yellow enzyme from Thermus scotoductus SA-01 UR - http://hdl.handle.net/11427/21606 ER - en_ZA


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