Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags

Wijasa, Teodora Stella; Sylvester, Marc; Brocke‐Ahmadinejad, Nahal; Kummer, Markus P.; Brosseron, Frederic; Gieselmann, Volkmar; Heneka, Michael T.

doi:10.1016/j.jneumeth.2017.08.005

Cited by 5 publications

(8 citation statements)

References 19 publications

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“…An optimized protocol for mass spectrometry‐based proteomics, using iodoTMT TM isobaric tag labeling was applied to identify synapto‐SNO protein modifications during physiological aging and along the AD trajectory. The performance of this protocol was comparable to our previous results (Wijasa et al ).…”

Section: Discussionsupporting

confidence: 90%

“…An optimized protocol for mass spectrometry-based proteomics, using iodoTMT TM isobaric tag labeling was applied to identify synapto-SNO protein modifications during physiological aging and along the AD trajectory. The performance of this protocol was comparable to our previous results (Wijasa et al 2017). S-nitrosylation is a fragile modification with fast turnover, although the SNO stability can vary between different proteins (Paige et al 2008).…”

Section: Discussionsupporting

confidence: 89%

“…Synaptosomes were purified as described previously (Wijasa et al ). In brief, synaptosomes were isolated from mouse and human brain samples and directly placed in a Teflon‐glass homogenizer with ninefold volume of tissue weight in ice‐cold 0.32 M sucrose (consists of 50 mM Tris‐Acetat, 1 mM EGTA, 1 mM EDTA, 1 mM AEBSF, cOmplete protease inhibitor cocktail 0.2% V/V, 5 mM Na 2 H 2 P 2 O 7 , 5 mM NaF, 2 mM NaVO 3 , pH 7.4).…”

Section: Methodsmentioning

confidence: 99%

“…All chemicals and antibodies used in this study are listed in detail in the supplementary file methods section. Isobaric mass tag labeling using iodoTMT™ (Thermo Fisher Scientific), nanocapillary high‐performance liquid chromatography tandem mass spectrometry, and basic mass spectrometry data processing were performed as previously described (Wijasa et al ). The identified proteins were annotated using Uniprot/SwissProt database (Bateman et al ) and then classified using Panther Gene Ontology (GO) (Thomas et al ) and Gorilla search engine (Eden et al ).…”

Section: Methodsmentioning

confidence: 99%

“…In this study, our aim was to characterize the NOS2‐dependent SNO of synaptic proteins as a fingerprint of neuroinflammation in mouse and human brain samples to discover new potential biomarker candidates for AD. Mass spectrometry‐based quantification with an irreversible Cys‐reactive reagent of Iodoacetyl Tandem Mass Tag (IodoTMT™, Thermo Fisher Scientific, Waltham, MA, USA) was used for both mice and human studies (Wijasa et al ). The age effect and genotype effect were analyzed in synaptosome samples prepared from wild‐type (WT) and AD‐transgenic mice.…”

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confidence: 99%

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Quantitative proteomics of synaptosome S‐nitrosylation in Alzheimer’s disease

Wijasa

Sylvester

Brocke‐Ahmadinejad

et al. 2019

Journal of Neurochemistry

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Increasing evidence suggests that both synaptic loss and neuroinflammation constitute early pathologic hallmarks of Alzheimer’s disease. A downstream event during inflammatory activation of microglia and astrocytes is the induction of nitric oxide synthase type 2, resulting in an increased release of nitric oxide and the post‐translational S‐nitrosylation of protein cysteine residues. Both early events, inflammation and synaptic dysfunction, could be connected if this excess nitrosylation occurs on synaptic proteins. In the long term, such changes could provide new insight into patho‐mechanisms as well as biomarker candidates from the early stages of disease progression. This study investigated S‐nitrosylation in synaptosomal proteins isolated from APP/PS1 model mice in comparison to wild type and NOS2−/− mice, as well as human control, mild cognitive impairment and Alzheimer’s disease brain tissues. Proteomics data were obtained using an established protocol utilizing an isobaric mass tag method, followed by nanocapillary high performance liquid chromatography tandem mass spectrometry. Statistical analysis identified the S‐nitrosylation sites most likely derived from an increase in nitric oxide (NO) in dependence of presence of AD pathology, age and the key enzyme NOS2. The resulting list of candidate proteins is discussed considering function, previous findings in the context of neurodegeneration, and the potential for further validation studies.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 89%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Quantitative proteomics of synaptosome S‐nitrosylation in Alzheimer’s disease

Wijasa

Sylvester

Brocke‐Ahmadinejad

et al. 2019

Journal of Neurochemistry

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Identification and Quantification of S-Sulfenylation Proteome of Mycobacterium tuberculosis under Oxidative Stress

Chen

Wang

et al. 2023

Microbiol Spectr

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Proteomic Analysis of S-Nitrosation Sites During Somatic Embryogenesis in Brazilian Pine, Araucaria angustifolia (Bertol.) Kuntze

et al. 2022

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Cysteine S-nitrosation is a redox-based post-translational modification that mediates nitric oxide (NO) regulation of various aspects of plant growth, development and stress responses. Despite its importance, studies exploring protein signaling pathways that are regulated by S-nitrosation during somatic embryogenesis have not been performed. In the present study, endogenous cysteine S-nitrosation site and S-nitrosated proteins were identified by iodo-TMT labeling during somatic embryogenesis in Brazilian pine, an endangered native conifer of South America. In addition, endogenous –S-nitrosothiol (SNO) levels and S-nitrosoglutathione reductase (GSNOR) activity were determined in cell lines with contrasting embryogenic potential. Overall, we identified an array of proteins associated with a large variety of biological processes and molecular functions with some of them already described as important for somatic embryogenesis (Class IV chitinase, pyruvate dehydrogenase E1 and dehydroascorbate reductase). In total, our S-nitrosoproteome analyses identified 18 endogenously S-nitrosated proteins and 50 in vitro S-nitrosated proteins (after GSNO treatment) during cell culture proliferation and embryo development. Furthermore, SNO levels and GSNOR activity were increased during embryo formation. These findings expand our understanding of the Brazilian pine proteome and shed novel insights into the potential use of pharmacological manipulation of NO levels by using NO inhibitors and donors during somatic embryogenesis.

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Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags

Abstract: Graphical abstract

Cited by 5 publications

References 19 publications

Quantitative proteomics of synaptosome S‐nitrosylation in Alzheimer’s disease

Quantitative proteomics of synaptosome S‐nitrosylation in Alzheimer’s disease

Identification and Quantification of S-Sulfenylation Proteome of Mycobacterium tuberculosis under Oxidative Stress

Proteomic Analysis of S-Nitrosation Sites During Somatic Embryogenesis in Brazilian Pine, Araucaria angustifolia (Bertol.) Kuntze

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