Dynamic posttranslational modifications of cytoskeletal proteins unveil hot spots under nitroxidative stress

Autor: Maria Fedorova, Andreia Mónico, Venukumar Vemula, Eva Griesser, Dolores Pérez-Sala
Přispěvatelé: German Research Foundation, European Commission, Agencia Estatal de Investigación (España), Ministerio de Economía y Competitividad (España), Instituto de Salud Carlos III, Mónico, Andreia, Pérez-Sala, Dolores, Mónico, Andreia [0000-0003-4855-8241], Pérez-Sala, Dolores [0000-0003-0600-665X]
Rok vydání: 2021
Předmět:
0301 basic medicine
Medicine (General)
Redox signaling
CID
collision-induced dissociation
Clinical Biochemistry
SIN-1
3-morpholinosydnonimine
Intermediate Filaments
Vimentin
Biochemistry
Nitroxidative stress
0302 clinical medicine
Tubulin
Biology (General)
Cytoskeleton
Intermediate filament
DOPA
dihydroxyphenylalanine
0303 health sciences
biology
ESI
electrospray ionization
Chemistry
Cell biology
LPP
lipid peroxidation products
Crosstalk (biology)
Signal transduction
Cellular model
actin
Research Paper
QH301-705.5
macromolecular substances
RNS
reactive nitrogen species
03 medical and health sciences
R5-920
ROS
reactive oxygen species
Microtubule
DDA
data dependent acquisition
Cysteine
Actin
030304 developmental biology
HNE
hydroxy-nonenal
HHE
hydroxyl-hexenal
MDA
malondialdehyde
PCA
principal component analysis
Organic Chemistry
DQ
dopaquinone
TMT
tandem mass tag
Cardiac cells
Lipoxidation
Cytoskeletal Proteins
030104 developmental biology
Posttranslational modifications
biology.protein
ETD
electron-transfer dissociation
PTM
posttranslational modification
Protein Processing
Post-Translational
030217 neurology & neurosurgery
Zdroj: Redox Biology
Digital.CSIC. Repositorio Institucional del CSIC
instname
Redox Biology, Vol 44, Iss, Pp 102014-(2021)
Digital.CSIC: Repositorio Institucional del CSIC
Consejo Superior de Investigaciones Científicas (CSIC)
ISSN: 2213-2317
Popis: 11 p.-2 fig.-3 tab.
The cytoskeleton is a supramolecular structure consisting of interacting protein networks that support cell dynamics in essential processes such as migration and division, as well as in responses to stress. Fast cytoskeletal remodeling is achieved with the participation of regulatory proteins and posttranslational modifications (PTMs).Redox-related PTMs are emerging as critical players in cytoskeletal regulation. Here we used a cellular model of mild nitroxidative stress in which a peroxynitrite donor induced transient changes in the organization of three key cytoskeletal proteins, i.e., vimentin, actin and tubulin. Nitroxidative stress-induced reconfiguration of intermediate filaments, microtubules and actin structures were further correlated with their PTM profiles and dynamics of the PTM landscape. Using high-resolution mass spectrometry, 62 different PTMs were identified and relatively quantified in vimentin, actin and tubulin, including 12 enzymatic, 13 oxidative and 2 nitric oxidederived modifications as well as 35 modifications by carbonylated lipid peroxidation products, thus evidencing the occurrence of a chain reaction with formation of numerous reactive species and activation of multiple signaling pathways. Our results unveil the presence of certain modifications under basal conditions and their modulation in response to stress in a target-, residue- and reactive species-dependent manner. Thus, some modifications accumulated during the experiment whereas others varied transiently. Moreover, we identified protein PTM “hot spots”, such as the single cysteine residue of vimentin, which was detected in seven modified forms, thus, supporting its role in PTM crosstalk and redox sensing. Finally, identification of novel PTMs in these proteins paves the way for unveiling new cytoskeleton regulatory mechanisms.
This work was funded by Deutsche Forschungsgemeinschaft (DFG;FE-1236/31 to M.F.), European Regional Development Fund (ERDF,European Union and Free State Saxony; 100146238 and 100121468 to MF), and MASSTRPLAN project funded by the Marie Sklodowska-Curie EU Framework for Research and Innovation Horizon 2020 (Grant Agreement No. 675132, to MF and DPS). EG STSM at CIB-CSIC was supported by COST Action CM1001. DPS work has been supported by Agencia Estatal de Investigación, MINECO/ERDF (grants SAF2015-68590R and RTI2018-097624-B-I00), and ISCIII/ERDF (RETIC ARADYAL RD16/0006/0021).
Databáze: OpenAIRE
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