Publication

SCA14 mutation V138E leads to partly unfolded PKCγ associated with an exposed C-terminus, altered kinetics, phosphorylation and enhanced insolubilization

Jezierska, J., Goedhart, J., Kampinga, H. H., Reits, E. A. & Verbeek, D. S., Mar-2014, In : Journal of Neurochemistry. 128, 5, p. 741-751 11 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Jezierska, J., Goedhart, J., Kampinga, H. H., Reits, E. A., & Verbeek, D. S. (2014). SCA14 mutation V138E leads to partly unfolded PKCγ associated with an exposed C-terminus, altered kinetics, phosphorylation and enhanced insolubilization. Journal of Neurochemistry, 128(5), 741-751. https://doi.org/10.1111/jnc.12491

Author

Jezierska, Justyna ; Goedhart, Joachim ; Kampinga, Harm H. ; Reits, Eric A. ; Verbeek, Dineke S. / SCA14 mutation V138E leads to partly unfolded PKCγ associated with an exposed C-terminus, altered kinetics, phosphorylation and enhanced insolubilization. In: Journal of Neurochemistry. 2014 ; Vol. 128, No. 5. pp. 741-751.

Harvard

Jezierska, J, Goedhart, J, Kampinga, HH, Reits, EA & Verbeek, DS 2014, 'SCA14 mutation V138E leads to partly unfolded PKCγ associated with an exposed C-terminus, altered kinetics, phosphorylation and enhanced insolubilization', Journal of Neurochemistry, vol. 128, no. 5, pp. 741-751. https://doi.org/10.1111/jnc.12491

Standard

SCA14 mutation V138E leads to partly unfolded PKCγ associated with an exposed C-terminus, altered kinetics, phosphorylation and enhanced insolubilization. / Jezierska, Justyna; Goedhart, Joachim; Kampinga, Harm H.; Reits, Eric A.; Verbeek, Dineke S.

In: Journal of Neurochemistry, Vol. 128, No. 5, 03.2014, p. 741-751.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Jezierska J, Goedhart J, Kampinga HH, Reits EA, Verbeek DS. SCA14 mutation V138E leads to partly unfolded PKCγ associated with an exposed C-terminus, altered kinetics, phosphorylation and enhanced insolubilization. Journal of Neurochemistry. 2014 Mar;128(5):741-751. https://doi.org/10.1111/jnc.12491


BibTeX

@article{44010af2e64546c0b49ce77e968e93c8,
title = "SCA14 mutation V138E leads to partly unfolded PKCγ associated with an exposed C-terminus, altered kinetics, phosphorylation and enhanced insolubilization",
abstract = "The protein kinase C γ (PKCγ) undergoes multistep activation and participates in various cellular processes in Purkinje cells. Perturbations in its phosphorylation state, conformation or localization can disrupt kinase signalling, such as in spinocerebellar ataxia type 14 (SCA14) that is caused by missense mutations in PRKCG encoding for PKCγ. We previously showed that SCA14 mutations enhance PKCγ membrane translocation upon stimulation owing to an altered protein conformation. As the faster translocation did not result in an increased function, we examined how SCA14 mutations induce this altered conformation of PKCγ and what the consequences of this conformational change are on PKCγ life cycle. Here, we show that SCA14-related PKCγ-V138E exhibits an exposed C-terminus as shown by fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy in living cells, indicative of its partial unfolding. This conformational change was associated with faster phorbol 12-myristate 13-acetate-induced translocation and accumulation of fully phosphorylated PKCγ in the insoluble fraction, which could be rescued by coexpressing PDK1 kinase that normally triggers PKCγ autophosphorylation. We propose that the SCA14 mutation V138E causes unfolding of the C1B domain and exposure of the C-terminus of the PKCγ-V138E molecule, resulting in a decrease of functional kinase in the soluble fraction. Here, we show that the mutation V138E of the protein kinase C γ (PKCγ) C1B domain (PKCγ-V138E), which is implicated in spinocerebellar ataxia type 14, exhibits a partially unfolded C-terminus. This leads to unusually fast phorbol 12-myristate 13-acetate-induced membrane translocation and accumulation of phosphorylated PKCγ-V138E in the insoluble fraction, causing loss of the functional kinase. In contrast to general chaperones, coexpression of PKCγ's 'natural chaperone', PDK1 kinase, could rescue the PKCγ-V138E phenotype.",
keywords = "C1B domain, conformational change, PDK1, protein kinase C, spinocerebellar ataxia, PROTEIN-KINASE-C, SPINOCEREBELLAR ATAXIA TYPE-14, PHOSPHOINOSITIDE-DEPENDENT KINASE, CEREBELLAR-ATAXIA, DOWN-REGULATION, BETA-II, ALPHA, MEMBRANE, DIACYLGLYCEROL, IDENTIFICATION",
author = "Justyna Jezierska and Joachim Goedhart and Kampinga, {Harm H.} and Reits, {Eric A.} and Verbeek, {Dineke S.}",
note = "{\textcopyright} 2013 International Society for Neurochemistry.",
year = "2014",
month = mar,
doi = "10.1111/jnc.12491",
language = "English",
volume = "128",
pages = "741--751",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Blackwell Publishing Ltd",
number = "5",

}

RIS

TY - JOUR

T1 - SCA14 mutation V138E leads to partly unfolded PKCγ associated with an exposed C-terminus, altered kinetics, phosphorylation and enhanced insolubilization

AU - Jezierska, Justyna

AU - Goedhart, Joachim

AU - Kampinga, Harm H.

AU - Reits, Eric A.

AU - Verbeek, Dineke S.

N1 - © 2013 International Society for Neurochemistry.

PY - 2014/3

Y1 - 2014/3

N2 - The protein kinase C γ (PKCγ) undergoes multistep activation and participates in various cellular processes in Purkinje cells. Perturbations in its phosphorylation state, conformation or localization can disrupt kinase signalling, such as in spinocerebellar ataxia type 14 (SCA14) that is caused by missense mutations in PRKCG encoding for PKCγ. We previously showed that SCA14 mutations enhance PKCγ membrane translocation upon stimulation owing to an altered protein conformation. As the faster translocation did not result in an increased function, we examined how SCA14 mutations induce this altered conformation of PKCγ and what the consequences of this conformational change are on PKCγ life cycle. Here, we show that SCA14-related PKCγ-V138E exhibits an exposed C-terminus as shown by fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy in living cells, indicative of its partial unfolding. This conformational change was associated with faster phorbol 12-myristate 13-acetate-induced translocation and accumulation of fully phosphorylated PKCγ in the insoluble fraction, which could be rescued by coexpressing PDK1 kinase that normally triggers PKCγ autophosphorylation. We propose that the SCA14 mutation V138E causes unfolding of the C1B domain and exposure of the C-terminus of the PKCγ-V138E molecule, resulting in a decrease of functional kinase in the soluble fraction. Here, we show that the mutation V138E of the protein kinase C γ (PKCγ) C1B domain (PKCγ-V138E), which is implicated in spinocerebellar ataxia type 14, exhibits a partially unfolded C-terminus. This leads to unusually fast phorbol 12-myristate 13-acetate-induced membrane translocation and accumulation of phosphorylated PKCγ-V138E in the insoluble fraction, causing loss of the functional kinase. In contrast to general chaperones, coexpression of PKCγ's 'natural chaperone', PDK1 kinase, could rescue the PKCγ-V138E phenotype.

AB - The protein kinase C γ (PKCγ) undergoes multistep activation and participates in various cellular processes in Purkinje cells. Perturbations in its phosphorylation state, conformation or localization can disrupt kinase signalling, such as in spinocerebellar ataxia type 14 (SCA14) that is caused by missense mutations in PRKCG encoding for PKCγ. We previously showed that SCA14 mutations enhance PKCγ membrane translocation upon stimulation owing to an altered protein conformation. As the faster translocation did not result in an increased function, we examined how SCA14 mutations induce this altered conformation of PKCγ and what the consequences of this conformational change are on PKCγ life cycle. Here, we show that SCA14-related PKCγ-V138E exhibits an exposed C-terminus as shown by fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy in living cells, indicative of its partial unfolding. This conformational change was associated with faster phorbol 12-myristate 13-acetate-induced translocation and accumulation of fully phosphorylated PKCγ in the insoluble fraction, which could be rescued by coexpressing PDK1 kinase that normally triggers PKCγ autophosphorylation. We propose that the SCA14 mutation V138E causes unfolding of the C1B domain and exposure of the C-terminus of the PKCγ-V138E molecule, resulting in a decrease of functional kinase in the soluble fraction. Here, we show that the mutation V138E of the protein kinase C γ (PKCγ) C1B domain (PKCγ-V138E), which is implicated in spinocerebellar ataxia type 14, exhibits a partially unfolded C-terminus. This leads to unusually fast phorbol 12-myristate 13-acetate-induced membrane translocation and accumulation of phosphorylated PKCγ-V138E in the insoluble fraction, causing loss of the functional kinase. In contrast to general chaperones, coexpression of PKCγ's 'natural chaperone', PDK1 kinase, could rescue the PKCγ-V138E phenotype.

KW - C1B domain

KW - conformational change

KW - PDK1

KW - protein kinase C

KW - spinocerebellar ataxia

KW - PROTEIN-KINASE-C

KW - SPINOCEREBELLAR ATAXIA TYPE-14

KW - PHOSPHOINOSITIDE-DEPENDENT KINASE

KW - CEREBELLAR-ATAXIA

KW - DOWN-REGULATION

KW - BETA-II

KW - ALPHA

KW - MEMBRANE

KW - DIACYLGLYCEROL

KW - IDENTIFICATION

U2 - 10.1111/jnc.12491

DO - 10.1111/jnc.12491

M3 - Article

C2 - 24134140

VL - 128

SP - 741

EP - 751

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 5

ER -

ID: 14138864