Insights into the importance of WPD-loop sequence for activity and structure in protein tyrosine phosphatases

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dc.creatorRuidan Shen
dc.creatorRory M. Crean
dc.creatorKeith J. Olsen
dc.creatorMarina Corbella Morató
dc.creatorAna Rita Calixto
dc.creatorTeisha Richan
dc.creatorTiago Antônio da Silva Brandão
dc.creatorRyan D. Berry
dc.creatorAlex Tolman
dc.creatorPatrick Loria
dc.creatorSean Johnson
dc.creatorShina Caroline Lynn Kamerlin
dc.creatorAlvan C. Hengge
dc.date.accessioned2023-10-19T19:44:47Z
dc.date.accessioned2025-09-08T22:51:56Z
dc.date.available2023-10-19T19:44:47Z
dc.date.issued2022
dc.description.sponsorshipOutra Agência
dc.format.mimetypepdf
dc.identifier.doihttps://doi.org/10.1039/D2SC04135A
dc.identifier.issn2041-6539
dc.identifier.urihttps://hdl.handle.net/1843/59751
dc.languageeng
dc.publisherUniversidade Federal de Minas Gerais
dc.relation.ispartofChemical Science
dc.rightsAcesso Aberto
dc.subjectProteinas - Análise
dc.subjectAnálise enzimática
dc.subjectAnálise conformacional
dc.subjectDinâmica molecular
dc.subjectCinética de enzimas
dc.subjectBioquímica
dc.subject.otherProtein tyrosine phosphatases (PTPs)
dc.subject.otherChimeric protein
dc.subject.otherConformational equilibria
dc.subject.otherMolecular dynamics (MD)
dc.subject.otherKinetic isotope effects (KIEs)
dc.titleInsights into the importance of WPD-loop sequence for activity and structure in protein tyrosine phosphatases
dc.typeArtigo de periódico
local.citation.epage13540
local.citation.issue45
local.citation.spage13524
local.citation.volume13
local.description.resumoProtein tyrosine phosphatases (PTPs) possess a conserved mobile catalytic loop, the WPD-loop, which brings an aspartic acid into the active site where it acts as an acid/base catalyst. Prior experimental and computational studies, focused on the human enzyme PTP1B and the PTP from Yersinia pestis, YopH, suggested that loop conformational dynamics are important in regulating both catalysis and evolvability. We have generated a chimeric protein in which the WPD-loop of YopH is transposed into PTP1B, and eight chimeras that systematically restored the loop sequence back to native PTP1B. Of these, four chimeras were soluble and were subjected to detailed biochemical and structural characterization, and a computational analysis of their WPD-loop dynamics. The chimeras maintain backbone structural integrity, with somewhat slower rates than either wild-type parent, and show differences in the pH dependency of catalysis, and changes in the effect of Mg2+. The chimeric proteins' WPD-loops differ significantly in their relative stability and rigidity. The time required for interconversion, coupled with electrostatic effects revealed by simulations, likely accounts for the activity differences between chimeras, and relative to the native enzymes. Our results further the understanding of connections between enzyme activity and the dynamics of catalytically important groups, particularly the effects of non-catalytic residues on key conformational equilibria.
local.identifier.orcidhttps://orcid.org/0000-0003-2670-1035
local.identifier.orcidhttps://orcid.org/0000-0003-4709-5353
local.identifier.orcidhttps://orcid.org/0000-0002-2248-9789
local.identifier.orcidhttps://orcid.org/0000-0001-9209-868X
local.identifier.orcidhttps://orcid.org/0000-0002-1123-0413
local.identifier.orcidhttps://orcid.org/0000-0001-5528-2293
local.identifier.orcidhttps://orcid.org/0000-0002-7783-3014
local.identifier.orcidhttps://orcid.org/0000-0002-0941-3560
local.identifier.orcidhttps://orcid.org/0000-0001-7992-2494
local.identifier.orcidhttps://orcid.org/0000-0002-3190-1173
local.identifier.orcidhttps://orcid.org/0000-0002-5696-2087
local.publisher.countryBrasil
local.publisher.departmentICX - DEPARTAMENTO DE QUÍMICA
local.publisher.initialsUFMG
local.url.externahttps://pubs.rsc.org/en/content/articlelanding/2022/SC/D2SC04135A

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