Protein-ribofuranosyl interactions activate orotidine 5′-monophosphate decarboxylase for catalysis

dc.creatorJudith R. Cristobal
dc.creatorTiago Antônio da Silva Brandão
dc.creatorArchie C. Reyes
dc.creatorJohn P. Richard
dc.date.accessioned2023-09-29T12:08:00Z
dc.date.accessioned2025-09-09T01:23:49Z
dc.date.available2023-09-29T12:08:00Z
dc.date.issued2021
dc.identifier.doihttps://doi.org/10.1021/acs.biochem.1c00589
dc.identifier.issn1520-4995
dc.identifier.urihttps://hdl.handle.net/1843/59026
dc.languageeng
dc.publisherUniversidade Federal de Minas Gerais
dc.relation.ispartofBiochemistry
dc.rightsAcesso Restrito
dc.subjectProteinas
dc.subjectAnálise enzimática
dc.subjectCinética de enzimas
dc.subjectMecanismos de reações orgânicas
dc.subjectReações químicas
dc.subjectBioquímica
dc.subject.otherChemical reactions
dc.subject.otherKinetic parameters
dc.subject.otherOrganic reactions
dc.subject.otherPeptides and proteins
dc.subject.otherStabilization
dc.titleProtein-ribofuranosyl interactions activate orotidine 5′-monophosphate decarboxylase for catalysis
dc.typeArtigo de periódico
local.citation.epage3373
local.citation.issue45
local.citation.spage3362
local.citation.volume60
local.description.resumoThe role of a global, substrate-driven, enzyme conformational change in enabling the extraordinarily large rate acceleration for orotidine 5′-monophosphate decarboxylase (OMPDC)-catalyzed decarboxylation of orotidine 5′-monophosphate(OMP) is examined in experiments that focus on the interactions between OMPDC and the ribosyl hydroxyl groups ofOMP. TheD37 and T100′side chains of OMPDC interact, respectively, with the C-3′and C-2′hydroxyl groups of enzyme-boundOMP.D37G and T100′A substitutions result in 1.4 kcal/mol increases in the activation barrier ΔG⧧for catalysis of decarboxylation of thephosphodianion-truncated substrate 1-(β-D-erythrofuranosyl) orotic acid (EO) but result in larger 2.1−2.9 kcal/mol increases in ΔG⧧for decarboxylation of OMPand for phosphite dianion-activated decarboxylation of EO. This shows that these substitutions reduce transition-state stabilization by the Q215, Y217, and R235 side chains at the dianion binding site. The D37G and T100′A substitutions result in <1.0 kcal/mol increases in ΔG⧧ for activation of OMPDC-catalyzed decarboxylation of the phosphoribofuranosyl-truncated substrate FO by phosphite dianions. Experiments to probe the effect of D37 and T100′substitutions on the kinetic parameters for D-glycerol 3-phosphate and D-erythritol 4-phosphate activators of OMPDC-catalyzed decarboxylation of FO show that ΔG⧧for sugar phosphate-activated reactions is increased byca.2.5 kcal/mol for each −OH interaction eliminated by D37G or T100′A substitutions. We conclude that the interactions between the D37 and T100′side chainsand ribosyl or ribosyl-like hydroxyl groups are utilized to activate OMPDC for catalysis of decarboxylation of OMP, EO, and FO.
local.identifier.orcidhttps://orcid.org/0000-0002-7783-3014
local.identifier.orcidhttps://orcid.org/0000-0001-9955-393X
local.identifier.orcidhttps://orcid.org/0000-0002-0440-2387
local.publisher.countryBrasil
local.publisher.departmentICX - DEPARTAMENTO DE QUÍMICA
local.publisher.initialsUFMG
local.url.externahttps://pubs.acs.org/doi/10.1021/acs.biochem.1c00589

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