Gel-based Protease Proteomics for Identifying the Novel Calpain Substrates in Dopaminergic Neuronal Cell

Kim, Chiho; Yun, Nuri; Lee, Young Mook; Jeong, Jae Yeong; Baek, Jeong Yeob; Song, Hoyoung; Ju, Chung; Youdim, Moussa B. H.; Jin, Byung Kwan; Kim, Won Ki; Oh, Young J.

doi:10.1074/jbc.m113.492876

Cited by 18 publications

(15 citation statements)

References 57 publications

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“…30 In accordance with the results of pharmacological inhibition of calpain, MN9D cells overexpressing calbindin-D-28K showed preservation of cdr2, resulting in more resistance to MPP + -induced cytotoxicity. Similarly, we demonstrated that MN9D cells overexpressing cdr2 are less vulnerable to MPP + -induced cytotoxic damage.…”

Section: Discussionsupporting

confidence: 75%

“…Consequently, pharmacological inhibition of these enzymes successfully blocked MPP + -induced degradation of cdr2 and subsequent dopaminergic neurodegeneration. Previous studies, including ones from our laboratory, report that MPP + -induced calpain activation via increased intracellular Ca 2+ leads to cleavage of numerous cellular substrates and contributes to neuronal cell death, 28, 30, 43, 44 supporting the notion that degradation of critical cellular proteins by activated calpain may be linked to dopaminergic neurodegeneration. An increase in intracellular free Ca 2+ is also known to occur in acute neurodegenerative conditions such as ischemic stroke and spinal cord injury.…”

Section: Discussionsupporting

confidence: 60%

“…MPP + -induced cell death is accompanied by or results from a burst of intracellular free Ca 2+ , leading to activation of calpain, a Ca 2+ -dependent cysteine protease in MN9D cells as previously demonstrated by us. 30, 37, 38 Therefore, we determined whether Ca 2+ -mediated calpain activation is responsible for MPP + -induced reduction of cdr2. As shown in Figure 5a, many MN9D cells stained positive for fluo-3, a Ca 2+ -sensitive fluorescent dye, 36 h after 50 μ M MPP + treatment.…”

Section: Resultsmentioning

confidence: 99%

“…18, 19, 20, 21, 22, 23, 24, 25, 26, 27 In our previous studies, we observed the activation of caspase and calpain in neurotoxin-induced dopaminergic neurodegeneration 28, 29 and found that degradation of endogenous substrates by activated proteases leads to neurodegeneration. 30, 31 Therefore, in the present study, we investigated the expression and protease-mediated regulation of cdr2 in experimental models of PD. We found that cdr2 is downregulated by calpain and the ubiquitin proteasome system and that the restoration of cdr2 levels renders dopaminergic neurons less vulnerable to 1-methyl-4-phenylpyridinium (MPP + )-mediated cytotoxicity.…”

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confidence: 99%

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Proteolytic degradation and potential role of onconeural protein cdr2 in neurodegeneration

Hwang

Lee

et al. 2016

Cell Death Dis

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Cerebellar degeneration-related protein 2 (cdr2) is expressed in the central nervous system, and its ectopic expression in tumor cells of patients with gynecological malignancies elicits immune responses by cdr2-specific autoantibodies and T lymphocytes, leading to neurological symptoms. However, little is known about the regulation and function of cdr2 in neurodegenerative diseases. Because we found that cdr2 is highly expressed in the midbrain, we investigated the role of cdr2 in experimental models of Parkinson's disease (PD). We found that cdr2 levels were significantly reduced after stereotaxic injection of 1-methyl-4-phenylpyridinium (MPP+) into the striatum. cdr2 levels were also decreased in the brains of post-mortem PD patients. Using primary cultures of mesencephalic neurons and MN9D cells, we confirmed that MPP+ reduces cdr2 in tyrosine hydroxylase-positive dopaminergic neuronal cells. The MPP+-induced decrease of cdr2 was primarily caused by calpain- and ubiquitin proteasome system-mediated degradation, and cotreatment with pharmacological inhibitors of these enzymes or overexpression of calcium-binding protein rendered cells less vulnerable to MPP+-mediated cytotoxicity. Consequently, overexpression of cdr2 rescued cells from MPP+-induced cytotoxicity, whereas knockdown of cdr2 accelerated toxicity. Collectively, our findings provide insights into the novel regulatory mechanism and potentially protective role of onconeural protein during dopaminergic neurodegeneration.

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Section: Discussionsupporting

confidence: 75%

Section: Discussionsupporting

confidence: 60%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Proteolytic degradation and potential role of onconeural protein cdr2 in neurodegeneration

Hwang

Lee

et al. 2016

Cell Death Dis

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“…These post-translational modifications are exponential due to a multitude of environmental effects spanning different individuals [6]. Magnitudes of proteomes span over 8-10 orders, whereas current mass spectrometer can only analyze up to 4 orders, add a challenge to this study [7].…”

Section: Introductionmentioning

confidence: 99%

Future of Urological Diagnosis Using Proteomics: A Review of Literature

Dagur¹,

Jason²,

hi³

et al. 2016

Transl Biomed

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Proteomics is a large-scale study of proteins. Several technologies are used to isolate and identify proteins of various spectra, which is useful in differentiating complexity of proteins. Critical role of proteomics can be applied to the field of urology, as a means of noninvasive biomarkers to diagnose patients with benign and malignant nephro-urological diseases. The accuracy of proteomic analysis findings in predicting specific clinical pathologies continues to draw controversy, thus further studies are necessary to confirm the diagnosis. Future of diagnosis in urology is expected to be done by proteomic urinalysis.

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The ubiquitin–proteasome system and autophagy mutually interact in neurotoxin‐induced dopaminergic cell death models of Parkinson's disease

Jang

Chung

2022

FEBS Letters

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Precise control of the two major proteolysis systems [i.e. ubiquitin-proteasome system (UPS) and autophagy-lysosomal pathway (ALP)] is important for proper cell function. Here, we explored whether UPS and ALP affect each other in two neurotoxin-based cell death models of Parkinson's disease. Monitoring UPS and ALP activity using their specific reporter plasmids revealed that treatment with the neurotoxin MPP + or the neurotoxin 6-OHDA decreased proteasome activity in dopaminergic MN9D cells. Interestingly, ALP inhibition relieved or potentiated the decrease in proteasome activity induced by the two toxins. Moreover, suppression of proteasome activity promoted 6-OHDA-induced excessive autophagic flux, potentiating ALP dysregulation. In contrast, MPP + -induced impairment of ALP was alleviated by proteasome inhibition. These findings suggest a dynamic interplay between UPS and ALP operating in MN9D cells under two distinct toxinmediated cell death pathways.

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Gel-based Protease Proteomics for Identifying the Novel Calpain Substrates in Dopaminergic Neuronal Cell

Cited by 18 publications

References 57 publications

Proteolytic degradation and potential role of onconeural protein cdr2 in neurodegeneration

Proteolytic degradation and potential role of onconeural protein cdr2 in neurodegeneration

Future of Urological Diagnosis Using Proteomics: A Review of Literature

The ubiquitin–proteasome system and autophagy mutually interact in neurotoxin‐induced dopaminergic cell death models of Parkinson's disease

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