Protein phosphorylation and protein phosphatasesDe Panne, Belgium, September 19–24, 1999

Zołnierowicz, Stanisław; Bollen, Mathieu

doi:10.1093/emboj/19.4.483

Cited by 169 publications

(138 citation statements)

References 31 publications

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“…This is in agreement with results showing that some PP1␣-targeting proteins have multiple sites of interaction with PP1 (56). We can conclude that caspase-9 is a PP1␣ substrate, and we entertain the possibility that it might also be a PP1␣-regulatory subunit.…”

Section: Discussionsupporting

confidence: 93%

Identification of PP1α as a Caspase-9 Regulator in IL-2 Deprivation-Induced Apoptosis

Dessauge

Cayla

Albar

et al. 2006

The Journal of Immunology

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One of the mechanisms that regulate cell death is the reversible phosphorylation of proteins. ERK/MAPK phosphorylates caspase-9 at Thr125, and this phosphorylation is crucial for caspase-9 inhibition. Until now, the phosphatase responsible for Thr125 dephosphorylation has not been described. Here, we demonstrate that in IL-2-proliferating cells, phosphorylated serine/threonine phosphatase type 1α (PP1α) associates with phosphorylated caspase-9. IL-2 deprivation induces PP1α dephosphorylation, which leads to its activation and, as a consequence, dephosphorylation and activation of caspase-9 and subsequent dissociation of both molecules. In cell-free systems supplemented with ATP caspase-9 activation is induced by addition of cytochrome c and we show that in this process PP1α is indispensable for triggering caspase-9 as well as caspase-3 cleavage and activation. Moreover, PP1α associates with caspase-9 in vitro and in vivo, suggesting that it is the phosphatase responsible for caspase-9 dephosphorylation and activation. Finally, we describe two novel phosphatase-binding sites different from the previously described PP1α consensus motifs, and we demonstrate that these novel sites mediate the interaction of PP1α with caspase-9.

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Section: Discussionsupporting

confidence: 93%

Identification of PP1α as a Caspase-9 Regulator in IL-2 Deprivation-Induced Apoptosis

Dessauge

Cayla

Albar

et al. 2006

The Journal of Immunology

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“…Phosphorylation, the most prevalent covalent modification of proteins in eukaryotic cells, affects approximately one-third of all proteins at any given time. 25 Phosphorylation is catalyzed by a variety of protein kinases including protein kinase A, one of the most well studied in cocaine abuse, 4 which phosphorylates serine and threonine residues on multiple targets including cAMP response element-binding protein. 26,27 Detecting changes in posttranslationally modified proteins in human postmortem tissue is difficult at best; however, similar assessments in nonhuman primate models of cocaine abuse are possible and have the advantage of recapitulating behavioral aspects of human drug intake and offering neuroanatomical and biochemical similarities compared with other species used as models.…”

Section: Introductionmentioning

confidence: 99%

Integrative proteomic analysis of the nucleus accumbens in rhesus monkeys following cocaine self-administration

2008

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The reinforcing effects and long-term consequences of cocaine self-administration have been associated with brain regions of the mesolimbic dopamine pathway, namely the nucleus accumbens (NAc). Studies of cocaine-induced biochemical adaptations in rodent models have advanced our knowledge; however, unbiased detailed assessments of intracellular alterations in the primate brain are scarce, yet essential, to develop a comprehensive understanding of cocaine addiction. To this end, two-dimensional difference in gel electrophoresis (2D-DIGE) was used to compare changes in cytosolic protein abundance in the NAc between rhesus monkeys selfadministering cocaine and controls. Following image normalization, spots with significantly differential image intensities (P < 0.05) were identified, excised, trypsin digested and analyzed by matrix-assisted laser-desorption ionization time-of-flight time-of-flight (MALDI-TOF-TOF). In total, 1098 spots were subjected to statistical analysis with 22 spots found to be differentially abundant of which 18 proteins were positively identified by mass spectrometry. In addition, approximately 1000 protein spots were constitutively expressed of which 21 proteins were positively identified by mass spectrometry. Increased levels of proteins in the cocaine-exposed monkeys include glial fibrillary acidic protein, syntaxin-binding protein 3, protein kinase C isoform, adenylate kinase isoenzyme 5 and mitochondrial-related proteins, whereas decreased levels of proteins included β-soluble N-ethylmaleimide-sensitive factor attachment protein and neural and non-neural enolase. Using a complimentary proteomics approach, the differential expression of phosphorylated proteins in the cytosolic fraction of these subjects was examined. Two-dimensional gel electrophoresis (2DGE) was followed by gel staining with Pro-Q Diamond phosphoprotein gel stain, enabling differentiation of approximately 150 phosphoprotein spots between the groups. Following excision and trypsin digestions, MALDI-TOF-TOF was used to confirm the identity of 15 cocaine-altered phosphoproteins. Significant increased levels were detected for γ-aminobutyric acid type A receptor-associated protein 1, 14-3-3 γ-protein, glutathione S-transferase and brain-type aldolase, whereas significant decreases were observed for β-actin, Rab GDP-dissociation inhibitor, guanine deaminase, peroxiredoxin 2 isoform b and Results from these studies indicate coordinated dysregulation of proteins related to cell structure, signaling, metabolism and mitochondrial function. These data extend and compliment previous studies of cocaine-induced biochemical alterations in human post-mortem brain tissue, using an animal model that closely recapitulates the human condition and provide new insight into the molecular basis of the disease and potential targets for pharmacotherapeutic intervention.

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“…About 25-30% of the proteins in eukaryotic cells are estimated to be phosphorylated at a certain content in the life cycle. Therefore, characterization of the phosphorylation is an important issue in proteomic analysis [4]. Numerous methods such as radioactive 32 P-labeling and detection [5], immunological methods [6], and Edman degradation [7] have been developed [8,9], although these techniques are relatively time consuming and laborious.…”

mentioning

confidence: 99%

Highly efficient and selective enrichment of phosphopeptides using porous anodic alumina membrane for MALDI-TOF MS analysis

Wang

Chen

et al. 2007

J. Am. Soc. Mass Spectrom.

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Because of its good biocompatibility, high surface-to-volume ratio, and distinct surface electrical properties, porous anodic alumina (PAA) membrane has been used to selectively enrich phosphopeptides from a mixture of synthetic peptides and tryptic digest product of ␤-casein by a direct MALDI-TOF MS analysis. As we reported previously, PAA membrane has strong incorporation ability to the phosphate anion. Herein, we describe the application of PAA membrane as a selective sampling absorbent for phosphopeptides. The PAA membrane could enrich phosphopeptides with high efficiency and selectivity; for example, the tryptic digest product of ␤-casein at a concentration as low as 4 ϫ 10 Ϫ9 M can be satisfactorily detected. Compared to that from the nonenriching peptide mixture, the MS signal of the phosphorylated peptides enriched by the PAA membrane is remarkably improved. In addition, acidic peptides have insignificant influence on the enriching process. Results show that the adsorption of phosphate anions on the PAA membrane plays a determining role in achieving highly selective enriching capacity toward phosphopeptides. The feasibility of PAA membranes as specific absorbents for phosphopeptides is also demonstrated. (J Am Soc Mass Spectrom 2007, 18, 1387-1395

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Protein phosphorylation and protein phosphatasesDe Panne, Belgium, September 19–24, 1999

Cited by 169 publications

References 31 publications

Identification of PP1α as a Caspase-9 Regulator in IL-2 Deprivation-Induced Apoptosis

Identification of PP1α as a Caspase-9 Regulator in IL-2 Deprivation-Induced Apoptosis

Integrative proteomic analysis of the nucleus accumbens in rhesus monkeys following cocaine self-administration

Highly efficient and selective enrichment of phosphopeptides using porous anodic alumina membrane for MALDI-TOF MS analysis

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