Glyceraldehyde-3-phosphate dehydrogenase: activity inhibition and protein overexpression in rotenone models for Parkinson's disease

Huang, Jinsha; Xiong, Nian; Chen, C.; Xiong, Jing; Jia, Min; Zhang, Z.; Cao, Xin; Liang, Zhihou; Sun, Shan; Lin, Zhicheng; Wang, T.

doi:10.1016/j.neuroscience.2011.06.050

Cited by 19 publications

(13 citation statements)

References 60 publications

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“…However, recent work has shown that deficiencies in GAPDH activity or expression are possible factors in neurodegenerative disorders such as Alzheimer’s and Parkinson’s Disease (PD) [2]. In the PD rotenone mouse model, GAPDH activity was overexpressed and activity was potently inhibited [3]. Several neurodegenerative diseases are characterized by the accumulation of protein aggregates [4].…”

Section: Introductionmentioning

confidence: 99%

Inactivation of glyceraldehyde-3-phosphate dehydrogenase by the dopamine metabolite, 3,4-dihydroxyphenylacetaldehyde

Vanle

Florang

Murry

et al. 2017

Biochemical and Biophysical Research Communications

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Background The aldehyde metabolite of dopamine, 3,4-dihydroxyphenylacetaldehyde (DOPAL) is an endogenous neurotoxin implicated in Parkinson’s Disease. Elucidating protein targets of DOPAL is essential in understanding it’s pathology. The enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a target of DOPAL. Methods GAPDH activity was measured via reduction of NAD+ cofactor (340 nm). Protein aggregation was assessed with SDS-PAGE methods and specific modification via chemical probes. Results Low micromolar levels of DOPAL caused extensive GAPDH aggregation and irreversibly inhibited enzyme activity. The inactivation of GAPDH was dependent on both the catechol and aldehyde moieties of DOPAL. It is suggested that Cys are modified and oxidized by DOPAL. Conclusions The mechanism by which DOPAL modifies GAPDH can serve as a mechanistic explanation to the pathological events in Parkinson’s Disease.

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Section: Introductionmentioning

confidence: 99%

Inactivation of glyceraldehyde-3-phosphate dehydrogenase by the dopamine metabolite, 3,4-dihydroxyphenylacetaldehyde

Vanle

Florang

Murry

et al. 2017

Biochemical and Biophysical Research Communications

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“…A study showed that GAPDH was the most extensively oxidized metabolic enzyme and activity was impaired in PD brains versus healthy brain samples 79 . Its activity and expression were reduced in a rotenone (PD cell model) 80 , and this correlates well with the fact that PD is often accompanied with impaired energetic metabolism pathways. Some reports claim that during high oxidative stress, GAPDH is oxidized and translocates to the nucleus, causing accumulation within the nucleus and activation of apoptosis via Caspase 3 81 .…”

Section: The Catecholaldehyde Hypothesis and Role Of 34-dihydroxyphesupporting

confidence: 57%

Reactive dopamine metabolites and neurotoxicity

Vanle¹

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I would like to thank them for their teaching and reviewing of my data at scientific conferences. A special thanks goes to Dr. Duffel whose lab has worked closely with ours; he always had time to discuss the 'mysteries of enzyme-drug interactions' with me, and the brainstorming of new ideas regarding the anomalies of enzyme assays. I would also like to thank Dr. Quinn for having office hours, and teaching one of the most challenging courses I had taken in graduate school-Mechanisms of Organic Reactions. I would also like to thank Dr. Kevin Rice for his active role in personally recruiting me and the other four classmates of Fall 2010. The idea of successfully finishing graduate school seems inconceivable without the camaraderie and help of my fellow classmates: Eric Rodriguez, Ioana Craciun, Aashay Shah, and Colin Higgins. We were truly a unified class with the same eagerness to learn and to advance science. Special thanks goes to Eric Rodriguez for the countless hours of scientific discussion and for the discovery of new solutions to the small and large problems that arose with instrumentation and cell assays. From over 2000 miles away, I would also like to acknowledge the unwavering and wholehearted support of my family and friends; especially my mother and father, Claudia and Hein, my grandmother Lola, Andrew and Christina. And last but not least, Bella and Zoe for being the best friends a woman could have. v ABSTRACT Parkinson's disease (PD) is a slow-progressive neurodegenerative disorder affecting 5-6 million people around the globe. The disease is manifested by the rapid deterioration of dopaminergic cells in the substantia nigra portion of the brain; however, the pathological mechanism of selective dopaminergic neuronal death is unknown. A reduction in levels of 3,4-dihydroxyphenylacetaldehyde (DOPAL) is biologically critical as this aldehyde has been shown to be toxic to dopaminergic cells and is a highly reactive electrophile. Investigating neuronal protein targets is essential in determining the cause of toxicity. An essential protein-GAPDH (e.g., glyceraldehyde-3-phosphate dehydrogenase) is an abundantly expressed enzyme known for its glycolytic activity, and recent research has implicated its role in oxidative stress-mediated neuronal death. This work positively shows GAPDH as a target for DOPAL modification, and, for the first time, DOPAL is identified as a potent inhibitor for GAPDH enzymatic activity. LC-MS and other chemical probes (ie. thiol and amine modifiers) show that DOPAL modifies specific amino acids: Lys, Arg, and modifies Cys and Met residues in the cofactor bindingdomain of GAPDH. The enzyme inhibition is also time and DOPAL dose-dependent. DOPAL has a unique structure, containing two reactive functional groups: an aldehyde and catechol ring. In-house syntheses of DOPAL analogues, containing the catechol group and lacking the aldehyde, and vice versa have been tested on GAPDH and do not inhibit or modify GAPDH. Therefore, both the catechol and aldehyde groups of DOPAL are specific to b...

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“…GAPDH is a critical housekeeping gene with significant implications in nuclear events and apoptosis. GAPDH-Siah-1interactions, provoked by cellular stress such as nitrosylation, has been shown to be involved in Parkinson's disease, AD (19), cerebral ischemia-reperfusion injury (20) and acute lung injury (21). Thus we decided to use our PLA based cell assay to investigate the effect of deprenyl, which has been shown to inhibit GAPDH and act as potential drug candidates for PD and other neurodegenerative disorders.…”

Section: Inhibition Of Gapdh-siah-1 Pla Using Deprenylmentioning

confidence: 99%

“…It has been shown that GAPDH binds to Siah-1 following S-nitrosylation of the catalytic cysteine of GAPDH, stabilizes otherwise unstable Siah-1, and GAPDH-Siah-1 complex translocates to the nucleus ( Figure 2) resulting in degradation of the nuclear targets of Siah-1 and subsequent enhancement of apoptosis (9). Thus, GAPDH is not only recognized as NO stress sensor but also GAPDH -Siah-1 complex is implicated in various diseases such as Alzheimer's disease (AD), Parkinson's disease (19), cerebral ischemia-reperfusion injury (20) , and acute lung injury (21). Interestingly, compounds (deprenyl and CGP-3466 [dibenzo-(b,f)oxepin-10ylmethyl-methyl-prop-2-ynylamine]) that inhibit GAPDH have been shown to act as potential drug candidates for PD and other neurodegenerative disorders, likely by disrupting the GAPDH -Siah-1 complex (11).…”

Section: Introductionmentioning

confidence: 99%

Glyceraldehyde-3-Phosphate Dehydrogenase and Seven in Absentia Homolog -1 Interactions in Mammalian Cells Visualized Using Proximity Ligation Assay

Singh¹,

Melm²

2016

Preprint

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Proteins seldom function in isolation and thus protein-protein interactions are critical in understanding the molecular basis of diseases and health (1,2). There are several well established techniques that are used to investigate protein-protein interactions(3). Most of the methods require some form of genetic modification of the target protein and thus always adds extra steps. However, Proximity Ligation Assay(4-6) (PLA) aka Duolink® is one such method that requires no genetic modification of the target protein and probes protein-protein interactions in fixed live cells and tissues. Briefly, PLA requires the use of primary antibodies specific to the proteins of interest. Once the sample (fixed cells or tissues) is incubated with species specific primary antibodies, secondary antibodies that are conjugated with oligonucleotides (also known as PLUS and MINUS probes respectively) and connecter oligonucleotides are added. This complex is ligated if the two PLUS and MINUS probes are within 40nm of each other. The resulting nucleic acid is amplified using rolling circle amplification and then probed with appropriate fluorescent probes. If the two proteins are interacting, one could visualize the interaction as a single red foci (for example Far Red Detection) using fluorescent microscopy.Here, we used PLA to probe protein-protein interactions between Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and seven in absentia homolog -1 (Siah-1)an E3 ubiquitin ligase.We first use PLA to show that GAPDH and Siah-1 proteins exist endogenously in the cytosol of multiple mammalian cell lines. Our data suggest the use of DU145 and T98G cell lines to show translocation of the GAPDH-Siah-1 complex. Next, we used common nitrosylation agents (7,8) (S-nitrosoglutathione-GSNO and S-Nitroso-N-acetyl-DL-pencillamine-SNAP) in different concentrations and observed that GAPDH and Siah-1 interact presumably due to the nitrosylation of the former, which is consistent with previous studies(9, 10). Interestingly, no interactions were observed between the two proteins in the absence of GSNO or SNAP indicating that nitrosylation might be critical for GAPDH-Siah1 interactions. Our results suggest that GAPDH-Siah-1 interactions originate in the cytosol and migrate to the nucleus under the conditions tested. We quantify the PLA signal using Duolink® Image Tool and observe a clear enhancement of GAPDH-Siah-1 PLA signal upon treating the cells with GSNO or SNAP. Next, we used R-(-)-Deprenyl (deprenyl), a known inhibitor of GAPDH 4 , and show that it abrogates GAPDH-Siah-1 PLA complex under the conditions tested. Finally, our data suggest that PLA can detect and quantify the GAPDH-Siah1 complex; a well-known protein-protein interaction implicated in neurodegeneration(9-11) and thus could be a method of choice for similar applications.

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Glyceraldehyde-3-phosphate dehydrogenase: activity inhibition and protein overexpression in rotenone models for Parkinson's disease

Cited by 19 publications

References 60 publications

Inactivation of glyceraldehyde-3-phosphate dehydrogenase by the dopamine metabolite, 3,4-dihydroxyphenylacetaldehyde

Inactivation of glyceraldehyde-3-phosphate dehydrogenase by the dopamine metabolite, 3,4-dihydroxyphenylacetaldehyde

Reactive dopamine metabolites and neurotoxicity

Glyceraldehyde-3-Phosphate Dehydrogenase and Seven in Absentia Homolog -1 Interactions in Mammalian Cells Visualized Using Proximity Ligation Assay

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