Weak domain stability and higher ca 2+ binding affinity contribute to allostery between the d/e linker and n-helix of cardiac troponin c

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dc.creatorMayra de Amorim Marques
dc.creatorAdolfo Henrique de Moraes Silva
dc.creatorJerson Lima da Silva
dc.creatorJose R. Pinto
dc.creatorGuilherme Augusto Piedade de Oliveira
dc.date.accessioned2022-10-06T23:43:36Z
dc.date.accessioned2025-09-08T22:57:30Z
dc.date.available2022-10-06T23:43:36Z
dc.date.issued2018-02-02
dc.format.mimetypepdf
dc.identifier.doihttps://doi.org/10.1016/j.bpj.2017.11.2336
dc.identifier.issn0006-3495
dc.identifier.urihttps://hdl.handle.net/1843/46067
dc.languageeng
dc.publisherUniversidade Federal de Minas Gerais
dc.relation.ispartofBiophysical Journal
dc.rightsAcesso Aberto
dc.subjectBatimento cardíaco
dc.subjectEnzimas alostéricas
dc.subjectMiocárdio
dc.subjectDoenças
dc.subjectMutação (Biologia)
dc.subject.otherHypertrophic cardiomyopathy (HCM)
dc.subject.otherSarcomeric proteins
dc.subject.otherCardiac dysfunction
dc.subject.otherCardiac troponin
dc.subject.otherProtein
dc.titleWeak domain stability and higher ca 2+ binding affinity contribute to allostery between the d/e linker and n-helix of cardiac troponin c
dc.typeArtigo de periódico
local.citation.epage422a
local.citation.issue3 Sup. 1
local.citation.spage421a
local.citation.volume114
local.description.resumoHypertrophic cardiomyopathy (HCM) is an inherited myopathy caused by the production of anomalous sarcomeric proteins that can lead to severe cardiac dysfunction. Here, we used structural and biophysical approaches to better understand the pathogenesis of a cardiac troponin C (cTnC) C84Y mutation located in the D/E linker, first reported in a 17-year-old proband, presenting with left-ventricular hypertrophy. Despite the relevance of HCM disease, little is known concerning the function of the D/E linker and allosteric phenomena governing cTnC Ca2+ affinity. Monitored by bis-ANS fluorescence, Ca2+-titrations reveal that C84Y exhibits enhanced Ca2+-binding affinity in both domains and conformational changes compared to WT. Although WT and C84Y display distinct Ca2+-binding behaviors, the overall dimensional values and molecular envelopes generated by small-angle-X-ray scattering data remains similar. Using circular-dichroism, C84Y revealed significantly lower thermostability in non-Ca2+-bound form compared to WT. Most of our understanding of the molecular mechanisms underlying how troponin and troponin peptides switch muscle contraction “on” and “off” has been derived using experimental NMR techniques. Currently, no experimental techniques are available that allow the understanding of protein regulatory/dynamic processes at the molecular level of large, multi-domain protein complexes. To further unravel molecular changes in C84Y, three-dimensional NMR experiments were performed for backbone assignment. The largest chemical shifts were observed in N-Helix residues and at the end of D-helix and D/E linker. NMR-derived backbone amide temperature-coefficients indicate different temperature-dependent conformational changes exist between WT and C84Y Carr-Purcell-Meiboom-Gill relaxation dispersion (CPMG-RD) and R1/R2 experiments were used to probe the population and exchanging rates of C84Y compared to WT. This work sought to elucidate: main structural components underlying this pathological mutation, novel allosteric mechanisms, and the role of D/E linker in cTnC.
local.identifier.orcidhttps://orcid.org/0000-0002-4131-4634
local.identifier.orcidhttps://orcid.org/0000-0002-3344-4084
local.identifier.orcidhttps://orcid.org/0000-0001-9523-9441
local.identifier.orcidhttps://orcid.org/0000-0001-9092-4976
local.identifier.orcidhttps://orcid.org/0000-0002-0063-5888
local.publisher.countryEstados unidos
local.publisher.departmentICX - DEPARTAMENTO DE QUÍMICA
local.publisher.initialsUFMG
local.url.externahttps://www.sciencedirect.com/science/article/pii/S0006349517335683?via%3Dihub

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