Repository logo
 
Publication

Closure of the human TKFC active site: comparison of the apoenzyme and the complexes formed with either triokinase or FMN cyclase substrates

dc.contributor.authorRodrigues, Joaquim Rui
dc.contributor.authorCameselle, José Carlos
dc.contributor.authorCabezas, Alicia
dc.contributor.authorRibeiro, João
dc.date.accessioned2019-05-14T11:28:09Z
dc.date.available2019-05-14T11:28:09Z
dc.date.issued2019-03-04
dc.description.abstractHuman triokinase/flavin mononucleotide (FMN) cyclase (hTKFC) catalyzes the adenosine triphosphate (ATP)-dependent phosphorylation of D-glyceraldehyde and dihydroxyacetone (DHA), and the cyclizing splitting of flavin adenine dinucleotide (FAD). hTKFC structural models are dimers of identical subunits, each with two domains, K and L, with an L2-K1-K2-L1 arrangement. Two active sites lie between L2-K1 and K2-L1, where triose binds K and ATP binds L, although the resulting ATP-to-triose distance is too large (≈14 Å) for phosphoryl transfer. A 75-ns trajectory of molecular dynamics shows considerable, but transient, ATP-to-DHA approximations in the L2-K1 site (4.83 Å or 4.16 Å). To confirm the trend towards site closure, and its relationship to kinase activity, apo-hTKFC, hTKFC:2DHA:2ATP and hTKFC:2FAD models were submitted to normal mode analysis. The trajectory of hTKFC:2DHA:2ATP was extended up to 160 ns, and 120-ns trajectories of apo-hTKFC and hTKFC:2FAD were simulated. The three systems were comparatively analyzed for equal lengths (120 ns) following the principles of essential dynamics, and by estimating site closure by distance measurements. The full trajectory of hTKFC:2DHA:2ATP was searched for in-line orientations and short distances of DHA hydroxymethyl oxygens to ATP γ-phosphorus. Full site closure was reached only in hTKFC:2DHA:2ATP, where conformations compatible with an associative phosphoryl transfer occurred in L2-K1 for significant trajectory time fractions.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.3390/ijms20051099pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.8/3953
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectTriokinasept_PT
dc.subjectDihydroxyacetone kinasept_PT
dc.subjectFMN cyclasept_PT
dc.subjectPhosphoryl transfer mechanismpt_PT
dc.subjectProtein domain mobilitypt_PT
dc.subjectActive-site closurept_PT
dc.subjectNormal mode analysispt_PT
dc.subjectMolecular dynamics simulationpt_PT
dc.subjectEssential dynamicspt_PT
dc.titleClosure of the human TKFC active site: comparison of the apoenzyme and the complexes formed with either triokinase or FMN cyclase substratespt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.issue5pt_PT
oaire.citation.startPage1099pt_PT
oaire.citation.titleInternational Journal of Molecular Sciencespt_PT
oaire.citation.volume20pt_PT
person.familyNameRodrigues
person.familyNameRibeiro
person.givenNameJoaquim Rui
person.givenNameJoão
person.identifier.ciencia-id9018-0B83-E2C6
person.identifier.orcid0000-0002-9756-1124
person.identifier.orcid0000-0002-6548-1402
person.identifier.ridL-4137-2014
person.identifier.ridK-6555-2014
person.identifier.scopus-author-id10242931100
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication52f6ffb2-43e9-4f78-b414-dbac46f80305
relation.isAuthorOfPublication36aa657e-2acc-4b21-8a64-a5221a17992f
relation.isAuthorOfPublication.latestForDiscovery52f6ffb2-43e9-4f78-b414-dbac46f80305

Files

Original bundle
Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
ijms-20-01099.pdf
Size:
5.83 MB
Format:
Adobe Portable Document Format
Description:
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.32 KB
Format:
Item-specific license agreed upon to submission
Description: